From lamastea from ipfw.edu Fri Jun 12 13:23:10 2009 From: lamastea from ipfw.edu (Arlis LaMaster) Date: Fri Jun 12 19:49:44 2009 Subject: [Microbiology] Kovac's reagent - difficulty making&In-Reply-To= Message-ID: <4A3264CE020000530006341A@gwia1.ipfw.edu> Hello Jeremy, Were you able to solve your problem making Kovac's reagent and it turning red? Ms. Arlis LaMaster lamastea@ipfw.edu From valentina_dafonseca from yahoo.com Wed Jun 17 08:57:49 2009 From: valentina_dafonseca from yahoo.com (valentina da fonseca) Date: Wed Jun 17 12:18:49 2009 Subject: [Microbiology] DNA Extraction Message-ID: <243130.23694.qm@web39101.mail.mud.yahoo.com> Dear all, I am working with tar sands ( oil sands from canada ) and I have been strugeling to extarct DNA out of my samples , I have been using the phenol chlorophorm extraction method but unfortunatelly with no possitive results. I am would like to ask anyone for some insight regarding otherr extraction methods to follow in order to extract DNA from my samples. Thank you val ____________________________________________________________________________________ ?Obt?n la mejor experiencia en la web! Descarga gratis el nuevo Internet Explorer 8. http://downloads.yahoo.com/ieak8/?l=e1 From mbarlett from microbio.umass.edu Thu Jun 18 12:26:23 2009 From: mbarlett from microbio.umass.edu (Melissa A. Barlett) Date: Thu Jun 18 13:43:07 2009 Subject: [Microbiology] Re: Microbio Digest, Vol 49, Issue 2 In-Reply-To: <200906181708.n5IH8kp28767@net.bio.net> References: <200906181708.n5IH8kp28767@net.bio.net> Message-ID: <4A3A78BF.1060105@microbio.umass.edu> I typically use kits for DNA extraction, with some additions, and have done well with difficult sediment samples. The two kits I tend to use are the Bio101 FastDNA SPIN kit for soil and the MoBio PowerSoil kit. In addition to these kits, I have on occasion added a heating step at 70C for 1 hour at the beginning while the cells are in the detergent buffer. Plant kits can also be good for removing other interfering substances. I have also had the problem where I got DNA, but not so much that it was easy to tell, however, the subsequent PCR was blocked by an interferent of some kind, making me think my extraction was bad. If you think this might be your problem, the Promega Wizard DNA clean-up kit is great for making bad DNA usable. Melissa University of Massachusetts > Message: 1 > Date: Wed, 17 Jun 2009 06:57:49 -0700 (PDT) > From: valentina da fonseca > Subject: [Microbiology] DNA Extraction > To: microbio@magpie.bio.indiana.edu > Message-ID: <243130.23694.qm@web39101.mail.mud.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Dear all, > I am working with tar sands ( oil sands from canada ) and I have been strugeling to extarct DNA out of my samples , I have been using the phenol chlorophorm extraction method but unfortunatelly with no possitive results. I am would like to ask anyone for some insight regarding otherr extraction methods to follow in order to extract DNA from my samples. > Thank you > val > > From medanrc from yahoo.com Mon Jun 22 06:34:33 2009 From: medanrc from yahoo.com (mohamed eda) Date: Mon Jun 22 09:45:31 2009 Subject: [Microbiology] glycerol stock for bacteria and fungi, Message-ID: <565448.43351.qm@web65514.mail.ac4.yahoo.com> Dear all hope you all fine and doing well I want to prepare a glycerol stock for bacteria and fungi, and I want to know if there are different techniques? for each, the second point is I did not have a -80 degrees freezer only -30 which I have, so it is possible to use it for the storage of the glycerol stock of the isolates and in this case is there is any special precaution in case of using -30 degree freezer. thanks in advance ???????????????????????????????? Yours Mohamed Eida Agricultural Microbiology Dept. National Research Center, Egypt Hiroshima University Graduate School of Biosphere Science Assessment of Environmental Dynamics Dept. Plant Environmental Science Lab. phone No. 090-6405-4222 (SoftBank) From rachnasingh.25 from gmail.com Sat Jun 27 21:28:57 2009 From: rachnasingh.25 from gmail.com (Rachna Singh) Date: Sat Jun 27 23:33:53 2009 Subject: [Microbiology] Re: Microbio Digest, Vol 49, Issue 5 In-Reply-To: <200906271705.n5RH5pp20475@net.bio.net> References: <200906271705.n5RH5pp20475@net.bio.net> Message-ID: <108188500906271928m323f9e16w7a196f3446469034@mail.gmail.com> Re: glycerol stock for bacteria and fungi, In our lab, we generally prepare glycerol stocks (bacteria) in brain-heart infusion (BHI) broth with 15 % glycerol and have stored the stocks at -20 as well as -80. Even we don't have access to -80 much. So, I generally use -20 stocks only. My -20 stocks, stored 3 years earlier, are working well - the bacteria are viable. Though the recommended temperature is -70 to -80, you can store at -20. On Sat, Jun 27, 2009 at 10:35 PM, wrote: > Send Microbio mailing list submissions to > microbio@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/microbio > or, via email, send a message with subject or body 'help' to > microbio-request@net.bio.net > > You can reach the person managing the list at > microbio-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Microbio digest..." > > > Today's Topics: > > 1. Re: glycerol stock for bacteria and fungi, (Bob) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 26 Jun 2009 16:03:06 -0700 > From: Bob > Subject: [Microbiology] Re: glycerol stock for bacteria and fungi, > To: microbio@net.bio.net > Message-ID: > Content-Type: text/plain; charset=ISO-8859-1 > > On Mon, 22 Jun 2009 04:34:33 -0700 (PDT), mohamed eda > wrote: > > >Dear all hope you all fine and doing well > >I want to prepare a glycerol stock for bacteria and fungi, and I want to > know if there are different techniques for each, > > > It is mainly empirical. If you have a procedure someone says works, > try it. Otherwise, you might try using 20% glycerol, with good healthy > cells. > > Run some tests to see if it works. Freeze, and thaw samples at > intervals; check viability. If it is pretty stable over a week, you're > probably ok. > > > >the second point is I did not have a -80 degrees freezer only -30 which I > have, so it is possible to use it for the storage of the glycerol stock of > the isolates and in this case is there is any special precaution in case of > using -30 degree freezer. > > > There are two concerns. > > One is simply that lower T is better -- things are slower. Probably > not a big deal. > > The other concern is whether the freezer holds T well. If samples are > stored near -20, T fluctuations can cause changes in ice structure; > not so good. > > If -30 is what you have, then try it. But you might also look into > maybe saving some samples elsewhere if -80 or liquid nitrogen are > available. Hey, storing some aliquots in at least a 2nd freezer is not > bad anyway -- just in case something goes wrong. > > > You might check with the ATCC. This is their business, and I suspect > they would share some info with you. > > bob > > > > ------------------------------ > > _______________________________________________ > Microbio mailing list > Microbio@net.bio.net > http://www.bio.net/biomail/listinfo/microbio > > End of Microbio Digest, Vol 49, Issue 5 > *************************************** >