Thursday, April 15, 2010
Fwd: What kits do people prefer for cDNA synth, and why?
---------- Forwarded message ----------
From: Nathan McCorkle <nmz787@gmail.com>
Date: Thu, Apr 15, 2010 at 1:35 PM
Subject: Re: What kits do people prefer for cDNA synth, and why?
To: diybio@googlegroups.com
The kit I used this morning included AMV reverse transcriptase, which has higher functioning temp which is better for denaturing secondary RNA structures... not sure how big a problem that is with mRNA.
From: Nathan McCorkle <nmz787@gmail.com>
Date: Thu, Apr 15, 2010 at 1:35 PM
Subject: Re: What kits do people prefer for cDNA synth, and why?
To: diybio@googlegroups.com
The kit I used this morning included AMV reverse transcriptase, which has higher functioning temp which is better for denaturing secondary RNA structures... not sure how big a problem that is with mRNA.
It doesn't seem like RNase H activity is much of a problem since I'm looking to amplify only a certain mRNA, rather than make a library... does that make sense, or is RNase H activity just bad/useless ?
--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
On Thu, Apr 15, 2010 at 2:07 PM, Stanley Han <zhan528@gmail.com> wrote:
NEB kit doesn't seem to be robust. I don't know whay. Invitrogen has a Rt enzyme that has no endo H activity so that yo get more cDNA.Clontech seems to be good too. They are all more expensive.
On Thu, Apr 15, 2010 at 12:19 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
--I am looking for cDNA synth kits and settled on this one at $4.33 a reaction:I was settled actually, until I used a promega kit this morning after doing a trizol-chloroform RNA extraction. My professor said that trizol is cheap and yields high amounts of RNA, and said that he uses the promega kit because our school prefers to order through VWR... I am still waiting to hear if we get a discount through VWR or not.If anyone has opinions about the kit I posted through NEB, or has a better recommendation please send quickly, as I'm trying to purchase stuff tomorrow, or at least have money set aside for its purchase.
--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/BioinformaticsYou received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
--
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
--
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
Fwd: Sequencer update
---------- Forwarded message ----------
From: TS Nowlan <apokruphos@gmail.com>
Date: Thu, Apr 15, 2010 at 1:20 PM
Subject: Re: Sequencer update
To: diybio@googlegroups.com
We have an ABI PRISM 310, an older model single-capillary sequencer. There is an electrical fault somewhere, which we are still in the process of hunting down / fixing. I'm not personally familiar with its materials (I'm used to just working with the data afterwards), but once we have it up and running I'm sure we'd happy to help test out your kit however we can.
-Tor
--
From: TS Nowlan <apokruphos@gmail.com>
Date: Thu, Apr 15, 2010 at 1:20 PM
Subject: Re: Sequencer update
To: diybio@googlegroups.com
We have an ABI PRISM 310, an older model single-capillary sequencer. There is an electrical fault somewhere, which we are still in the process of hunting down / fixing. I'm not personally familiar with its materials (I'm used to just working with the data afterwards), but once we have it up and running I'm sure we'd happy to help test out your kit however we can.
-Tor
On Thu, Apr 15, 2010 at 11:12 AM, Stanley Han <zhan528@gmail.com> wrote:
What type of ABI instrument do you have? I wonder if you will be interested in testing my dye removal kit with your instrument if you use capillery sequencer---PureSEQ?
On Thu, Apr 15, 2010 at 2:10 PM, Stanley Han <zhan528@gmail.com> wrote:
You need a enzyme that incorporate ddNTP efficiently for sequencing not regular Taq. Bigdye use this kind of Taq.
On Wed, Apr 14, 2010 at 7:39 PM, Mackenzie Cowell <mac@diybio.org> wrote:
I'm organizing a PCR genotyping workshop next weekend and have been looking for a diybio-friendly sanger sequencing provider (haven't actually made accounts or taked to anyone at any companies or sequencing centers yet, though). It would be extra cool to do some of the sequencing with diybio! If I got reagents, could you guys do some sequencing on one or two samples?Also, a general question about sanger sequencing: I'm probably going to use Taq, but I might purchase a higher-fidelity / higher-processivity polymerase. What sort of read lengths could I typically get from a commercial sanger sequencing provider? Like, 700-1000 bp? I would be interested in amplifying like 5kb, if possible.Cheers,Mac--
On Wed, Apr 14, 2010 at 7:23 PM, Bryan Bishop <kanzure@gmail.com> wrote:
---------- Forwarded message ----------
From: TS Nowlan <apokruphos@gmail.com>
Date: Wed, Apr 14, 2010 at 6:14 PM
Subject: Sequencer update
To: socal-diybio@googlegroups.com
Hey folks,
Things are moving slowly forward with our lovely sequencer, but I
thought it good to put all the docs and whatnot we've managed to find
together in one place. I've found a protocol for using it, a Windows
installer for ABI sequence analysis software (good to convert it into
a non-proprietary format for us to work with) and user's manuals for
the diagnostics system and the sequencer overall. I added links to all
that to the Lab Protocol wiki page, though I imagine it should have
its own page at some point. I'll likely be adding more this afternoon
as I find it.
Here's the link again for those that may have forgotten, or not made
accounts yet:
http://omegahelm.com/diybio/tiki-index.php?page=Lab+Protocols
And of course feel free to edit and add more stuff as you like.
-Tor
--
"And if ye cannot be saints of knowledge, then be at least its warriors."
-- Friedrich Nietzsche
--
- Bryan
http://heybryan.org/
1 512 203 0507
--
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
+1.231.313.9062 / mac@diybio.org / @100ideas--
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
--
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
"And if ye cannot be saints of knowledge, then be at least its warriors."
-- Friedrich Nietzsche
--
-- Friedrich Nietzsche
--
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
Subscribe to:
Posts (Atom)
Posts
