From owner-rapd@net.bio.net Fri Sep 04 23:00:00 1998 Path: biosci!daresbury!daresbury!server5.netnews.ja.net!server6.netnews.ja.net!nntp.news.xara.net!xara.net!dispose.news.demon.net!demon!woodstock.news.demon.net!demon!news.vienna.good.net!news.good.net!news.megsinet.net!uunet!in3.uu.net!server-b.cs.interbusiness.it!news.tin.it!not-for-mail From: excell@bware.it Newsgroups: bionet.molbio.rapd Subject: Announce: how to SEND EMAIL AND NEWS from your GSM PHONE Date: 26 Aug 1998 10:49:45 GMT Organization: eXcell GSM/SMS gateway Message-ID: <6s0p89$ifv$5148@nslave1.tin.it> NNTP-Posting-Host: rub182.mi03.mi.interbusiness.it Mime-Version: 1.0 Content-Type: text/plain Lines: 27 Your GSM phone is already connected to the Internet! Now you can send EMAIL and post to newsgroups directly from your GSM phone. Just write a short message (SMS) with the syntax: EMAIL user@host message and then send it to the "eXcell" phone number at +393388641732 If your phone does not have the @ character you can use the ! , like: EMAIL user!host message There are several options available, such as a personal signature, a personal email address configuration for getting back replies, personal mailing lists, anti-spamming, carbon copy, phone number anonymizer, etc.etc. The only cost for you is the SMS shipping; normally a few pennies. Our service is currently being used by the GSM users of 40 different countries all around the world, using more than 80 different GSM networks. And it will certainly work for you also, so give it a try! To know more about eXcell, and to get the complete user's guide, come and visit http://www.bware.it/excell ( no advertising banners ) Be an eXcellent GSM user! From owner-rapd@net.bio.net Fri Sep 04 23:00:00 1998 Path: biosci!LANET.COM.AR!lmcriado From: lmcriado@LANET.COM.AR ("liliana y manuel") Newsgroups: bionet.molbio.rapd Subject: Subscribe Date: 5 Sep 1998 05:52:21 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 33 Sender: daemon@net.bio.net Distribution: world Message-ID: <000801bdd8cf$26260e20$23e50dd1@default> NNTP-Posting-Host: net.bio.net This is a multi-part message in MIME format. ------=_NextPart_000_0005_01BDD8B5.E75F3AE0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable I would like to subscribe. E-mail: lmcriado@lanet.com.ar ------=_NextPart_000_0005_01BDD8B5.E75F3AE0 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable

I would like to = subscribe.

E-mail:=20 lmcriado@lanet.com.ar

------=_NextPart_000_0005_01BDD8B5.E75F3AE0-- From owner-rapd@net.bio.net Sat Sep 05 23:00:00 1998 Path: biosci!EUnet.yu!gdrinic From: gdrinic@EUnet.yu ("Goran Drinic") Newsgroups: bionet.molbio.rapd Subject: (none) Date: 6 Sep 1998 04:17:32 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <000001bdd987$a6c9d760$f2c6f7c2@Goran.EUnet.yu> NNTP-Posting-Host: net.bio.net SUBSCRIBE RAPD to biosci-server@net.bio.net From owner-rapd@net.bio.net Mon Sep 07 23:00:00 1998 Path: biosci!agate!newsfeed.berkeley.edu!howland.erols.net!vixen.cso.uiuc.edu!not-for-mail From: Jason Newsgroups: bionet.molbio.rapd Subject: testing. Date: Mon, 07 Sep 1998 19:30:21 -0700 Organization: University of Illinois at Urbana-Champaign Lines: 2 Message-ID: <35F496BD.9A095F89@uiuc.edu> NNTP-Posting-Host: spider.cso.uiuc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.06 [en] (Win95; I) X-Proxy-Client: kama@uiuc.edu from ppp43-ts3.pdnt.com From owner-rapd@net.bio.net Mon Sep 07 23:00:00 1998 Path: biosci!agate!newsfeed.berkeley.edu!howland.erols.net!vixen.cso.uiuc.edu!not-for-mail From: Jason Newsgroups: bionet.molbio.rapd Subject: testing. Date: Mon, 07 Sep 1998 19:30:50 -0700 Organization: University of Illinois at Urbana-Champaign Lines: 2 Message-ID: <35F496DA.1432B61A@uiuc.edu> NNTP-Posting-Host: spider.cso.uiuc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.06 [en] (Win95; I) X-Proxy-Client: kama@uiuc.edu from ppp43-ts3.pdnt.com testing. From owner-rapd@net.bio.net Sun Sep 13 23:00:00 1998 Path: biosci!daresbury!uninett.no!hugin.hsh.no!hotmail.com From: 19F.4Voyeur.@hotmail.com Newsgroups: bionet.molbio.rapd Subject: 19 F and topless;-) - i ve got a webcamera - ME ! Date: Sunday, 13 Sep 1998 10:31:23 -0600 Organization: http://www.afterhoursclub.com/club/cam/mystrip.htm Lines: 7 Message-ID: <13099810.3123@hotmail.com> NNTP-Posting-Host: ole.uninett.no X-Trace: snipp.uninett.no 905734043 27209 158.37.8.10 (14 Sep 1998 00:47:23 GMT) X-Complaints-To: news-abuse@uninett.no Cache-Post-Path: ole.uninett.no!unknown@hugin.hsh.no hi, my name is caolinei m a 19years french female student i just put an on line webcamera in my room i m seeking 4 voyeur to cyber ( i m bi) http://www.afterhoursclub.com/club/cam/mystrip.htm i m not shy.. and shaved, amateur only please!!!! if no serious please do not contact have a nice weekend HIFN5)5i>%AG From owner-rapd@net.bio.net Wed Sep 16 23:00:00 1998 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.rapd Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 17 Sep 1998 02:00:12 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 233 Sender: daemon@net.bio.net Distribution: world Message-ID: <199809170900.CAA17886@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. From owner-rapd@net.bio.net Wed Sep 16 23:00:00 1998 Path: biosci!news.stanford.edu!newsfeed.concentric.net!su-news-hub1.bbnplanet.com!news.bbnplanet.com!logbridge.uoregon.edu!infeed.is.co.za!feeder.is.co.za!news.mikom.csir.co.za!not-for-mail From: Michael Crampton Newsgroups: bionet.molbio.rapd Subject: Cloning of pcr fragment Date: Wed, 16 Sep 1998 12:01:38 +0200 Organization: Mikomtek/CSIR Lines: 3 Message-ID: <35FF8C81.E123DA83@csir.co.za> NNTP-Posting-Host: 146.64.120.62 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.02 [en] (Win95; I) I am looking for a protocol that allows the cloning of very small pcr fragments (75-200bp) into a t-tailed vector. From owner-rapd@net.bio.net Wed Sep 16 23:00:00 1998 Path: biosci!agate!newsfeed.berkeley.edu!news.maxwell.syr.edu!cpk-news-hub1.bbnplanet.com!atl-news-feed1.bbnplanet.com!news.bbnplanet.com!ox.mc.edu!not-for-mail From: Robert Hamilton Newsgroups: bionet.molbio.rapd Subject: Re: Cloning of pcr fragment Date: Thu, 17 Sep 1998 09:10:34 -0500 Organization: Mississippi College Lines: 6 Message-ID: <3601185A.4186@mc.edu> References: <35FF8C81.E123DA83@csir.co.za> Reply-To: rhamilto@mc.edu NNTP-Posting-Host: rhamilto.mc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0Gold (Win95; I; 16bit) Michael Crampton wrote: > > I am looking for a protocol that allows the cloning of very small pcr > fragments (75-200bp) into a t-tailed vector. Invitrogen Topo-TA cloning kit. From owner-rapd@net.bio.net Thu Sep 17 23:00:00 1998 Path: biosci!news.stanford.edu!newsfeed.berkeley.edu!news.maxwell.syr.edu!cpk-news-hub1.bbnplanet.com!atl-news-feed1.bbnplanet.com!news.bbnplanet.com!ox.mc.edu!not-for-mail From: Robert Hamilton Newsgroups: bionet.molbio.rapd Subject: Re: non-Mendelian AFLP bands; fresh vs frozen tissue Date: Fri, 18 Sep 1998 10:34:01 -0500 Organization: Mississippi College Lines: 2 Message-ID: <36027D69.41E8@mc.edu> References: Reply-To: rhamilto@mc.edu NNTP-Posting-Host: rhamilto.mc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0Gold (Win95; I; 16bit) Can't recombination create and eliminate such bands. What is the frequency of these bands among all bands? From owner-rapd@net.bio.net Thu Sep 17 23:00:00 1998 Path: biosci!news.stanford.edu!su-news-feed2.bbnplanet.com!su-news-hub1.bbnplanet.com!atl-news-feed1.bbnplanet.com!news.bbnplanet.com!ox.mc.edu!not-for-mail From: Robert Hamilton Newsgroups: bionet.molbio.rapd Subject: Re: non-Mendelian AFLP bands; fresh vs frozen tissue Date: Fri, 18 Sep 1998 14:20:28 -0500 Organization: Mississippi College Lines: 26 Message-ID: <3602B27C.32B5@mc.edu> References: <36027D69.41E8@mc.edu> Reply-To: rhamilto@mc.edu NNTP-Posting-Host: rhamilto.mc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0Gold (Win95; I; 16bit) Diana Wolf wrote: > What do you mean by 'recombination'? I don't think there is any > recombination in mitosis > I must have missed something, as I thought the essence of the problem in your original post was as follows: "When performing AFLP analysis on parents and offspring, I noticed a) bands that were present in both parents were absent in all (10) of their offspring; b) bands that were present in the offspring but absent their parents."...copy and paste from your original post. You then looked at frozen vs fresh tissue of the same type, and got a similar pattern, assuming that the phenomenon was due to freezing, but can come up with no explanation. Still, however, your original problem was bands in one generation, not in another, which leads to the possible effect of recombination...what is the effect of recombination on such analyses? The DNAeasy kits supposedly shears the DNA into 1000bp (approx.) fragments as I recall. I could see degradation eliminating fragments, but producing them? Perhaps there is an effect on the ends of the DNA fragments? I use RAPDs. Next time someone who uses AFLPs includes the running down of RAPDs in a talk, I will ask them about this particular problem :^) From owner-rapd@net.bio.net Thu Sep 17 23:00:00 1998 Path: biosci!agate!newsfeed.berkeley.edu!howland.erols.net!vixen.cso.uiuc.edu!news.indiana.edu!willow.bio.indiana.edu!user From: dewolf@bio.indiana.edu (Diana Wolf) Newsgroups: bionet.molbio.rapd Subject: Re: non-Mendelian AFLP bands; fresh vs frozen tissue Date: Fri, 18 Sep 1998 17:53:04 -0600 Organization: Indiana University Lines: 72 Message-ID: References: <36027D69.41E8@mc.edu> <3602B27C.32B5@mc.edu> NNTP-Posting-Host: willow.bio.indiana.edu In article <3602B27C.32B5@mc.edu>, rhamilto@mc.edu wrote: > Diana Wolf wrote: > > > What do you mean by 'recombination'? I don't think there is any > > recombination in mitosis > > > I must have missed something, as I thought the essence of the problem in > your original post was as follows: > > "When performing AFLP analysis on parents and offspring, I noticed a) > bands that were present in both parents were absent in all (10) of their > offspring; b) bands that were present in the offspring but absent their > parents."...copy and paste from your original post. you are right - I didn't think of that - I am too fixated on my freezing idea. I could have two seperate issues (recomb. eliminating the parental band or producing new bands in the offspring and then the fresh vs. frozen stuff). However, I guess I didn't tell the full story - in the parental I had a band that was present in males and absent in females. I looked and males and females in two full-sib families (the two families had different parents), and the band was present in 90% of males and absent in 90% of females. I then looked at two more full-sib families and found that the band was absent in all of the offspring. I think recomb. would only destroy the bands in a small percentage of progeny in each family (granted, my sample sizes are small - 5 males per family). > > You then looked at frozen vs fresh tissue of the same type, and got a the fresh and frozen tissues were collected from the same individual > similar pattern, assuming that the phenomenon was due to freezing, but > can come up with no explanation. Still, however, your original problem > was bands in one generation, not in another, which leads to the possible > effect of recombination...what is the effect of recombination on such > analyses? recombination between a '+' allele and a '-' might create two '-' alleles or even a completely novel allele. However the largest fragments on my gel are ..umm.. I can't find the sheet that says how big the bands on my ladder are - but they shouldn't be more than 1kb (prob more like 500bp max), and recombination within such a small region should be quite infrequent. > > The DNAeasy kits supposedly shears the DNA into 1000bp (approx.) > fragments as I recall. I could see degradation eliminating fragments, > but producing them? seems highly unlikely to me too >Perhaps there is an effect on the ends of the DNA > fragments? I use RAPDs. Next time someone who uses AFLPs includes the > running down of RAPDs in a talk, I will ask them about this particular > problem :^) hmm.. yes I think all markers are problematic - my experience hasn't shown AFLPs to be any more reproducable than RAPDs (I used to use those). However there is that paper that used AFLPs, RAPDs and something else (RFLPs or microsats) in a bunch of different labs, and compared the results to see if the gels looked the same. They found RAPDs difficult to reproduce, only had one band that didn't appear in all the labs when using AFLPs. Unfortunately...my experience is different... If you're interested, I'll find the reference for you (but maybe you'd rather ignore it ;-) Thanks tons for your ideas! Diana -- Biology Dept Indiana University From owner-rapd@net.bio.net Thu Sep 17 23:00:00 1998 Path: biosci!news.stanford.edu!su-news-feed2.bbnplanet.com!su-news-hub1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!netnews.com!newsfeed.enteract.com!news-xfer.siscom.net!streamer1.cleveland.iagnet.net!NewsNG.Chicago.Qual.Net!news.indiana.edu!pumpkin.bio.indiana.edu!user From: dewolf@bio.indiana.edu (Diana Wolf) Newsgroups: bionet.molbio.rapd Subject: non-Mendelian AFLP bands; fresh vs frozen tissue Date: Thu, 17 Sep 1998 22:26:41 -0600 Organization: Indiana University Lines: 68 Message-ID: NNTP-Posting-Host: pumpkin.bio.indiana.edu Hi, I'm having problems with non-Mendelian AFLP bands and don't know what to do. Any suggestions of causes or solutions will be greatly appreciated! I extracted DNA from full-sib families: I put healthy leaf tissue into coin envelopes and froze the tissue at -80C - then extracted the DNA 1-4 months later using the DNeasy plant extraction kit from Qiagen. I extracted DNA from the parents: I used healthy,fresh leaf tissue, and the DNeasy kit. DNA concentrations were measured with a florometer, and 300ng of DNA were used in each digest. When performing AFLP analysis on parents and offspring, I noticed a) bands that were present in both parents were absent in all (10) of their offspring; b) bands that were present in the offspring but absent their parents. I thought that this might have been due to the fact that offspring tissue was frozen, while parental tissue was fresh. Thus, I froze some parental tissue, extracted it (DNeasy), and did AFLP analysis on both the fresh and frozen extractions (fresh and frozen leaf samples were both from the same individuals, and both were in the same preamplification reaction and selective amplification reaction). The samples were analyzed with two different primer pairs. With one primer pair, I found 5 bands that are present in all frozen samples and absent in all fresh samples. With the other primer pair, I found three bands thare are present in all frozen samples and absent in the fresh sample; and one (short) band that was darker in the fresh samples. I don't know what is causing this, but I have a few ideas: a) Partial digestions are occurring in either fresh or frozen samples due to dirty (proteins) DNA. To test this, I eluquicked both fresh and frozed DNA and re-AFLP ed. The differences between fresh and frozed tissue remained, so dirty DNA can't be the problem. b) frozen tissue is degraded I ran whole-genomic DNA from fresh and frozen tissue on an agarose gel, and both looked the same. I only saw big clumps of large-sized DNA, no smear of smaller DNA c) One or both of the endonucleases (EcoR I and Mse I) may be methylation sensitive - and there may be methylation differences in fresh and frozen tissues (either due to freezing or because tissues were collected at different times). I e-mailed NEB to ask if they are methylation sensitive, however I don't know why there should be differences in methylation. d)freezing the tissue may help break up cell walls, causing DNA from frozen tissue to be enriched for organellar DNA. This doesn't explain the bands that are present in (fresh) parental tissue and absent in all of the (frozen) offspring. Any ideas or help will be greatly appreciated!! Thank you very much! Diana Wolf Biology Dept. Indiana University -- Diana Wolf Biology Department Indiana University From owner-rapd@net.bio.net Thu Sep 17 23:00:00 1998 Path: biosci!daresbury!not-for-mail From: "MTT E-mail Cimtar" Newsgroups: bionet.molbio.rapd Subject: [28742137812130061219] UJ BEJEGYZES az MTT E-mail Cimtar rendszereben Date: 18 Sep 1998 05:27:13 +0100 Lines: 60 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <6tsnf1$14i@mserv1.dl.ac.uk> Original-To: rapd@dl.ac.uk MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7BIT Kedves felhaszn�l�! Megtal�ltuk az �n e-mail c�m�t az Interneten. Ha szeretn�, hogy adatait az MTT E-mail C�mt�r rendszere tartalmazza, k�rj�k l�togassa meg szerver�nket a http://e_search.mtt.hu/ c�men. Ott tov�bbi adatokat adhat meg mag�r�l, �gy k�nnyebben tal�lj�k meg azok akik keresik. 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With kind regards, The MTT E-mail Directory Team From owner-rapd@net.bio.net Thu Sep 17 23:00:00 1998 Path: biosci!news.stanford.edu!newsfeed.berkeley.edu!newsfeed.enteract.com!chicago-news-feed1.bbnplanet.com!news.bbnplanet.com!web.onecall.net!news.indiana.edu!willow.bio.indiana.edu!user From: dewolf@bio.indiana.edu (Diana Wolf) Newsgroups: bionet.molbio.rapd Subject: Re: non-Mendelian AFLP bands; fresh vs frozen tissue Date: Fri, 18 Sep 1998 11:52:23 -0600 Organization: Indiana University Lines: 18 Message-ID: References: <36027D69.41E8@mc.edu> NNTP-Posting-Host: willow.bio.indiana.edu With primer pair 1: there are 18 monomorphic bands, 11 polymorphic bands, and 3 bands that are present in all frozen samples and absent in all fresh samples (bands that show this pattern are not included in "polymorphic bands") With primer pair 2: there are 10 monomorphic bands, 10 polymorphic bands, and if you count really pale bands, 9 bands that show the pattern. What do you mean by 'recombination'? I don't think there is any recombination in mitosis Diana In article <36027D69.41E8@mc.edu>, rhamilto@mc.edu wrote: > Can't recombination create and eliminate such bands. What is the > frequency of these bands among all bands? From owner-rapd@net.bio.net Fri Sep 18 23:00:00 1998 Path: biosci!TVI.IS!loy96 From: loy96@TVI.IS (Kraftworks) Newsgroups: bionet.molbio.rapd Subject: PREMIUM TV Channels.......No Monthly Bills ! Date: 19 Sep 1998 05:38:06 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 178 Sender: daemon@net.bio.net Distribution: world Message-ID: <199809192292CAA44988@funprojects.teleco.ulpgc.es> NNTP-Posting-Host: net.bio.net This is really cool! PREMIUM CHANNELS........Descrambled! EASY to assemble plans for only $6.00 ! YOU WILL BE WATCHING all your favorite pay stations featuring MOVIES, SPORTS. Adult entertainment, and any other scrambled signal NEXT WEEK! 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Learn how easy descrambling is! $ 6.00 for plans only $10.00 for variable capacitor only $16.00 for The easy to assemble plans and one variable capacitor! Pay by check or money order payable to: Kraftworks P.O. Box 11752 San Rafael, Ca. 94912 WE pay postage and handling! Please allow 10 days for delivery. The Mailing List that you are being mailed from was filtered against the Global Remove List. If you wish to be included in this REMOVE LIST please go to: http://remove-list.com It is a widely used list honored by all responsible emailers. Once on this list you will see a great reduction in advertisements. Thank You +....No Mo From owner-rapd@net.bio.net Sun Sep 20 23:00:00 1998 Path: biosci!TUSK.EDU!prakash From: prakash@TUSK.EDU ("C. S. Prakash") Newsgroups: bionet.molbio.rapd Subject: $11 MILLION NSF GRANT FOR MAIZE GENOME AT MISSOURI Date: 20 Sep 1998 18:22:29 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 83 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net (Looks like first of many US universities to reap the plant genome windfall!........Prakash) MU AWARDED $11 MILLION GRANT FROM THE NATIONAL SCIENCE FOUNDATION TO CREATE MAIZE RESEARCH CENTER ON CAMPUS September 17, 1998 University of Missouri Press Release COLUMBIA, Mo. -- On the University of Missouri-Columbia campus today, Chancellor Richard Wallace announced an $11 million grant from the National Science Foundation. The largest single research grant ever awarded to the University, it will create a center for maize genome research on campus. The NSF also awarded more than $500,000 to MU to enhance soybean research. "We are extremely pleased and proud that the National Science Foundation has recognized the University of Missouri-Columbia, and especially our researchers Ed Coe and Joe Polacco, national leaders in these crucial fields of research," Wallace said. "These grants are a major component for our life sciences strategy as MU works to overcome the challenges that await us in the 21st century." MU competed with more than 60 other institutions for the grants, of which 10 were awarded. The NSF grant was made possible because of an initiative, sponsored by Sen. Christopher "Kit" Bond, to place $40 million in the NSF budget for a competitive program on plant genomics related to economically important crops. "I am very excited that through a competitive process, the University has earned this significant grant, which will help put MU on the ground floor as we move forward into the new millennium that scientists are calling the 'century of biotechnology,'"Bond said. "This is a great achievement for the University of Missouri and represents another critical step forward in making Missouri the Silicon Valley of biotechnology." "We applaud Sen. Bond's visionary leadership on a program that will have major implications on feeding the world in the 21st century," said Jack Burns, vice provost of research. "This was a very selective process by the National Science Foundation and our researchers worked very hard to win the award. This grant underscores why life sciences is a key part of MU's mission enhancement proposal and will couple with the additional state resources to make MU a national and international leader in the area of plant science research." Part of the research will include enhancing the content and effectiveness of the maize genome database started by the U.S. Department of Agriculture in 1991. Other researchers will look at gene selection and it will be the responsibility of MU to link all of the information together in the databases. These resources will provide for much greater efficiency in mapping and identifying the 50,000 to 80,000 genes of maize. "Once we have the resources, knowledge and database complete, the possibilities are endless," Coe said. "As we learn more about how the systems of these plants work, we will know what we can manipulate. Benefits of this research include better yields, reduced fertilizer requirements and better quality food. The end result is a better quality of life not only for us, but for the entire planet." For the soybean research, scientists will attempt to identify and determine what each of the 50,000 to 100,000 soybean genes actually do in the plant. Once identified, scientists will know which genes to target for increased yield, and disease and pest resistance varieties. This work also will provide the scientific community with a large database filled with information that will increase the efficiency of current soybean research. MU scientists involved in the maize project will collaborate with scientists at Clemson University and the University of Georgia to develop the physical resources, information systems and student training. In the soybean project, MU is one of four university partners with Iowa State University, the University of Minnesota and the University of Illinois. The maize grant is for five years and the soybean grant is for four years. Both will go toward a national Center for Crop Genomics on the MU campus, which will be housed in the proposed life sciences buildi ************************************************ C. S. Prakash Tuskegee University Center for Plant Biotechnology Research Tuskegee, AL 36088, USA mailto:Prakash@tusk.edu http://agriculture.tusk.edu/biotech/biotech.html Phone (334) 727 8023; Fax (334) 727 8067 ************************************************ From owner-rapd@net.bio.net Sun Sep 20 23:00:00 1998 Path: biosci!news.stanford.edu!su-news-feed2.bbnplanet.com!su-news-hub1.bbnplanet.com!atl-news-feed1.bbnplanet.com!news.bbnplanet.com!ox.mc.edu!not-for-mail From: Robert Hamilton Newsgroups: bionet.molbio.rapd Subject: Re: non-Mendelian AFLP bands; fresh vs frozen tissue Date: Sun, 20 Sep 1998 19:46:41 -0500 Organization: Mississippi College Lines: 15 Message-ID: <3605A1F1.1527@mc.edu> References: <36027D69.41E8@mc.edu> <3602B27C.32B5@mc.edu> Reply-To: rhamilto@mc.edu NNTP-Posting-Host: rhamilto.mc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0Gold (Win95; I; 16bit) Diana Wolf wrote: > If you're interested, I'll find the reference for you (but maybe you'd > rather ignore it ;-) > One of the problems all such techniques have is "reproduceability in different labs, not so much due to the technique, I think, but the technicians! We get great RAPDS on an SSCP type gel. I like AFLPs, but they are far to expensive for me! If you ever find the solution to your problem, I would like to know it, it could be relevant to a lot of things! Thanks. Rob.