From yuxia.jiao from Vanderbilt.Edu Wed May 13 12:47:46 2009 From: yuxia.jiao from Vanderbilt.Edu (Jiao, Yuxia) Date: Wed May 13 13:14:35 2009 Subject: [Automated-sequencing] Chromas 1.44 Message-ID: <4A3D91BF57295A41B445D41A2E49EBBF0113E189@mailbe01.mc.vanderbilt.edu> Hi, there, I can't download the Chromas 1.44. Every time I clicked the link (http://www.technelysium.com.au/chromas14x.html),it came back with "Windows can't display the webpage" . do you have another link that I try? Thank you very much Yuxia From valet from iib.unsam.edu.ar Wed May 13 14:09:30 2009 From: valet from iib.unsam.edu.ar (Valeria Tekiel) Date: Wed May 13 15:19:34 2009 Subject: [Automated-sequencing] Chromas 1.44 In-Reply-To: <4A3D91BF57295A41B445D41A2E49EBBF0113E189@mailbe01.mc.vanderbilt.edu> References: <4A3D91BF57295A41B445D41A2E49EBBF0113E189@mailbe01.mc.vanderbilt.edu> Message-ID: Try with FinchTV, it is almost the same V. 2009/5/13 Jiao, Yuxia > Hi, there, > > > > I can't download the Chromas 1.44. Every time I clicked the link > (http://www.technelysium.com.au/chromas14x.html),it came back with > "Windows can't display the webpage" . do you have another link that I > try? > > > > Thank you very much > > > > Yuxia > > > > _______________________________________________ > Autoseq mailing list > Autoseq@net.bio.net > http://www.bio.net/biomail/listinfo/autoseq > > -- ________________________________________ Dra. Valeria Tekiel Instituto de Investigaciones Biotecnologicas, UNSAM-Conicet Av. Gral Paz 5445, Predio INTI; Edificio 19 B1650 KNA San Martin, Buenos Aires. Argentina TE : 5411 4580 7255-57 (int: 332); FAX: 5411 4752 9639 E. mail: valet@iib.unsam.edu.ar From dna0001 from flinders.edu.au Tue May 19 09:14:33 2009 From: dna0001 from flinders.edu.au (dna0001@flinders.edu.au) Date: Tue May 19 09:37:10 2009 Subject: [Automated-sequencing] injection solution which reduces rfu Message-ID: <1242742473.4a12bec9391eb@imp.flinders.edu.au> Hi all I work in a core lab setting, and have recently upgraded my Applied Biosystems 3100 Genetic Analyser to a 3130xl running POP-7. A client wants to perform fragment analysis with multiplexed PCR products, but often the peaks of interest are off-scale (i.e. > 7000 rfu). To the best of my knowledge, Gene Mapper v4.0 and Peak Scanner v1.0 will not analyse these samples due to the off-scale peaks. I've tried adding 0.1mM EDTA to the samples (we generally use HiDi formamide only for injection), but have found that this can either increase or decrease signal strength. Can anyone suggest a solution which will decrease signal strength? Will low melting point agarose achieve this? Any suggestions would be appreciated. Many thanks Oliver From dreynold from aecom.yu.edu Tue May 19 18:11:09 2009 From: dreynold from aecom.yu.edu (David Reynolds) Date: Tue May 19 22:45:14 2009 Subject: [Automated-sequencing] Re: Autoseq Digest, Vol 38, Issue 2 In-Reply-To: <200905191703.n4JH3Jp15719@net.bio.net> References: <200905191703.n4JH3Jp15719@net.bio.net> Message-ID: <20090519231059.AFD2E5E@post.aecom.yu.edu> Hi Oliver, Instead of, or in conjuction with an additive you can dilute the fragments to get reasonable RFUs. On our 3730 the PCR reactions were typically diluted 40 to 120 fold. We would titrate a few samples per marker at 40, 80 and 120 fold, then pick the best dilution for the rest of the study. Regards, Dave At 01:03 PM 5/19/2009, you wrote: >Send Autoseq mailing list submissions to > autoseq@net.bio.net > >To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/autoseq >or, via email, send a message with subject or body 'help' to > autoseq-request@net.bio.net > >You can reach the person managing the list at > autoseq-owner@net.bio.net > >When replying, please edit your Subject line so it is more specific >than "Re: Contents of Autoseq digest..." > > >Today's Topics: > > 1. injection solution which reduces rfu (dna0001@flinders.edu.au) > > >---------------------------------------------------------------------- > >Message: 1 >Date: Tue, 19 May 2009 23:44:33 +0930 >From: dna0001@flinders.edu.au >Subject: [Automated-sequencing] injection solution which reduces rfu >To: autoseq@magpie.bio.indiana.edu >Message-ID: <1242742473.4a12bec9391eb@imp.flinders.edu.au> >Content-Type: text/plain; charset=ISO-8859-1 > > > >Hi all > I work in a core lab setting, and have recently upgraded my Applied >Biosystems 3100 Genetic Analyser to a 3130xl running POP-7. A client wants to >perform fragment analysis with multiplexed PCR products, but often >the peaks of >interest are off-scale (i.e. > 7000 rfu). To the best of my knowledge, Gene >Mapper v4.0 and Peak Scanner v1.0 will not analyse these samples due to the >off-scale peaks. I've tried adding 0.1mM EDTA to the samples (we generally use >HiDi formamide only for injection), but have found that this can either >increase or decrease signal strength. Can anyone suggest a solution which will >decrease signal strength? Will low melting point agarose achieve this? Any >suggestions would be appreciated. > >Many thanks > >Oliver > > > >------------------------------ > >_______________________________________________ >Autoseq mailing list >Autoseq@net.bio.net >http://www.bio.net/biomail/listinfo/autoseq > >End of Autoseq Digest, Vol 38, Issue 2 >************************************** ________________________________________________________________ David Reynolds Director Genomics & Genetics Core Albert Einstein College of Medicine Price Center for Genetic and Translational Medicine Room 419 1301 Morris Park Avenue Bronx, New York 10461-1602 Tel: (718) 678-1157 Lab: (718) 678-1160 Fax: (718) 678-1016 E-mail: dreynold@aecom.yu.edu http://www.aecom.yu.edu/dna/ ________________________________________________________________ From pmiguel from purdue.edu Wed May 20 06:06:41 2009 From: pmiguel from purdue.edu (Phillip San Miguel) Date: Wed May 20 06:12:15 2009 Subject: [Automated-sequencing] Re: injection solution which reduces rfu In-Reply-To: References: Message-ID: dna0001@flinders.edu.au wrote: > > Hi all > I work in a core lab setting, and have recently upgraded my Applied > Biosystems 3100 Genetic Analyser to a 3130xl running POP-7. A client wants to > perform fragment analysis with multiplexed PCR products, but often the peaks of > interest are off-scale (i.e. > 7000 rfu). To the best of my knowledge, Gene > Mapper v4.0 and Peak Scanner v1.0 will not analyse these samples due to the > off-scale peaks. I've tried adding 0.1mM EDTA to the samples (we generally use > HiDi formamide only for injection), but have found that this can either > increase or decrease signal strength. Can anyone suggest a solution which will > decrease signal strength? Will low melting point agarose achieve this? Any > suggestions would be appreciated. > > Many thanks > > Oliver > Hi Oliver, You could try reducing the injection time. Or ask your customer to load less sample. Seems like adding salt (like EDTA) just competes with product such that larger fragments diminish in intensity, but smaller ones stay about the same. Phillip