From CHAVENS from uwyo.edu Fri Aug 1 12:13:13 2008 From: CHAVENS from uwyo.edu (Christopher M Havens) Date: Fri Aug 1 16:59:50 2008 Subject: [Microbiology] Candida culturing Message-ID: <6BE35B6278F7344684D6138B8379BB090F0420@TELEGRAPH3.uwyo.edu> Hello, I have been having some trouble maintaining cultures of candida which I have aquired from ATCC. My first plate, made from the rehydrated powder they send always looks fine, giving reasonable numbers of good sized, unifrom colonies. But when I make a culture from that plate, spin down the cells, and then dilute and make new plates for counting I always end up with plates overgrown with tiny colonies which look quite different from the colonies yielded from the original rehydration. The numbers of these small colonies gives improbably high counts given my dilutions. I feel quite confident that I am using good sterile technique and that my problem is not due to contamination. Additionally I can isolate any of these colonies, stain them, and they appear as candida when viewed microscopically. Does anyone know what I am experiencing? Is this due to spore formation? Any help would be really appreciated. --CH From kdevik from gmail.com Tue Aug 5 04:17:33 2008 From: kdevik from gmail.com (Kanchanadevi k) Date: Tue Aug 5 11:07:47 2008 Subject: [Microbiology] need help-how to make bacterial mass inoculum Message-ID: <8b3d015d0808050217u44815f22k9a564478ad9fbf15@mail.gmail.com> hi can you please tell me how to make bacterial culture in tub. actually, i will have to raise 5ltr of bacterial culture mass inoculum (Nutrient Broth) in a tub for field application. so, if you have any idea about this please share with me Thanks a lot. K.Kanchana Devi Research Scholar From farrlarr from isu.edu Tue Aug 5 11:28:04 2008 From: farrlarr from isu.edu (Larry Farrell) Date: Tue Aug 5 12:21:32 2008 Subject: [Microbiology] Re: need help-how to make bacterial mass inoculum In-Reply-To: References: Message-ID: <41957$48987f9b$4660@news.teranews.com> Kanchanadevi k wrote: > hi > > can you please tell me how to make bacterial culture in tub. > actually, i will have to raise 5ltr of bacterial culture mass > inoculum (Nutrient Broth) in a tub for field application. > so, if you have any idea about this > please share with me > > Thanks a lot. > > > > > > K.Kanchana Devi > Research Scholar Why does it have to be in a tub? If you are growing only 5 liters of culture, it would make much more sense to grow several smaller cultures (500 ml each in 1 liter flasks; 1 liter each in 2 liter flasks; etc.). It certainly isn't going to be easy to sterilize a tub, particularly given that most tubs available these days aren't made of materials that will withstand autoclave temperature. Even if the tub is made of such material, it will be difficult to prepare a cover that will adequately protect the medium inside from contamination once it is removed from the autoclave (and I don't want to be around when you are trying to remove the tub/medium from the autoclave, or transport it to the field, without spilling it all over). On the other hand, it is very easy to autoclave/move/inoculate 1 liter or 2 liter flasks (or even larger ones, if you have them). -- Larry D. Farrell, Ph.D. Professor Emeritus of Microbiology Idaho State University ** Posted from http://www.teranews.com ** From egillock from fhsu.edu Wed Aug 6 13:30:55 2008 From: egillock from fhsu.edu (egillock@fhsu.edu) Date: Wed Aug 6 17:04:14 2008 Subject: [Microbiology] need help-how to make bacterial mass inoculum Message-ID: If you need this large volume for some reason, you might consider using an autoclavable carboy with a magnetic stir bar for aeration and mixing. This large volume will be difficult to handle though. I agree with the others, your best bet would be a series of smaller flasks in a shaking incubator. ----------------------------------------------- Eric T. Gillock, Ph.D. Associate Professor Department of Biological Sciences Fort Hays State University Hays, KS 67601 ----------------------------------------------- From vasu.nanobiotech91 from gmail.com Fri Aug 8 04:09:29 2008 From: vasu.nanobiotech91 from gmail.com (shrinivas Dengeti) Date: Fri Aug 8 11:14:52 2008 Subject: [Microbiology] Suggestion for carrying out experiments with the genes involved in alkalophily of Bacillus sp. Message-ID: I have planned to carry out characterization of genes involved in alkalophilic nature of thermoalkalophilic Bacillus sp. Somebody can suggest me some of the methodology involved in PCR amplification of the genes or preliminary experiments required to understand molecular basis behind the nature. D. Shrinivas Research Scholar From vasu.nanobiotech91 from gmail.com Mon Aug 11 10:20:09 2008 From: vasu.nanobiotech91 from gmail.com (shrinivas Dengeti) Date: Mon Aug 11 11:15:41 2008 Subject: [Microbiology] Re: Suggestion for carrying out experiments with the genes involved in alkalophily of Bacillus sp. References: Message-ID: <611b2a84-5f22-468f-9f2f-5af9cd203c2c@o40g2000prn.googlegroups.com> On Aug 9, 7:03?am, Bob wrote: > On Fri, 8 Aug 2008 02:09:29 -0700 (PDT), shrinivas Dengeti > > wrote: > >I have planned to carry out characterization of genes involved in > >alkalophilic nature of thermoalkalophilic Bacillus sp. > >Somebody can suggest me some of the methodology involved in PCR > >amplification of the genes or preliminary experiments required to > >understand molecular basis behind the nature. > > What do you already know? Do you have a genome sequence? Do you know > genes of interest? What do you know about them? Predicted protein > sequences often offer clues about function. Do you have an expression > system for Bacillus genes? > > bob i have different gene sequence of the xylanase from the Bacillus sp. and i have PCR amplified the gene..but not yet sequenced. I want to know whether tranport protein play any role in the thermoalkalophilic nature or anyother factor responsible for it....if anyone workin on the aspect.please help me in this matter... with regards D.shrinivas From n.mvenge from imperial.ac.uk Thu Aug 14 07:39:00 2008 From: n.mvenge from imperial.ac.uk (Mvenge, Nyasha) Date: Thu Aug 14 09:37:41 2008 Subject: [Microbiology] Hygromycin marker for Non tuberculous mycobacteria Message-ID: <1CA1A859632B1F4989CB23F90D0A7829042C33B9@icex4.ic.ac.uk> I am writting to ask about what concentrations you have found to be effective when selecting for transformants of Non tuberculous mycobacteria such as M.avium,abscessus and chelonae. Currently we have been using 50 and 100 ucl/ml but we keep getting colonies growing effectively rendering selection a complete waste of time.Apparently selection should work at 50ucl/ml.Do you have any suggestions for any other antibiotic apart from kanamycin? Thank you Grace. From vasu.nanobiotech91 from gmail.com Sat Aug 16 07:31:47 2008 From: vasu.nanobiotech91 from gmail.com (shrinivas Dengeti) Date: Sat Aug 16 16:36:01 2008 Subject: [Microbiology] Re: Suggestion for carrying out experiments with the genes involved in alkalophily of Bacillus sp. References: <611b2a84-5f22-468f-9f2f-5af9cd203c2c@o40g2000prn.googlegroups.com> Message-ID: <23c2c070-c5f7-4021-bcf5-a87babef203b@z72g2000hsb.googlegroups.com> thank you bob.. for showing such a interest in helpin me out. n thank you for the link too. hey u still didnt got me..actually to say..i want to know whether the transport involved in maintaining the alkalophilic nature of bacillus for ex. ATPase , Na/H antiporyter pump,.. and one more thing do they play any role in expresion of the enzyme thank yoyu With regards. D.shrinivas Bob wrote: > On Mon, 11 Aug 2008 08:20:09 -0700 (PDT), shrinivas Dengeti > wrote: > > > > >i have different gene sequence of the xylanase from the Bacillus sp. > >and i have PCR amplified the gene..but not yet sequenced. > >I want to know whether tranport protein play any role in the > >thermoalkalophilic nature or anyother factor responsible for it....if > >anyone workin on the aspect.please help me in this matter... > > > > A couple of quick thoughts. You've probably thought of these points > yourself, so I?ll be brief. > > It is interesting to have an extracellular enzyme from an alkalophile. > One can compare its properties with a "normal" relative, and then look > for an explanation in the 3D structure. Do you have a good 3D > structure for any of the related enzymes? > > Of course, the general alkalophilic nature of the bug probably lies > largely in keeping the cytoplasm near "normal". > > > Do you know about the following papers: > > http://springerlink.metapress.com/content/h3m1703x3711215t/?p=1bd8409162894aaa99d979a1fe61676d&pi=5 > > http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V24-4JRM055-1&_user=4420&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_version=1&_urlVersion=0&_userid=4420&md5=6c95e774252e769ea34297f1bc792e49 > > I bet you already have them :-) -- but figured it safer to ask, to be > sure. If you don't have access, feel free to email me for copies, at > my yahoo acct: b_bruner > > regards, > > bob