Check RTPrimerDB (http://medgen.ugent.be/rtprimerdb), PrimerBank (http://pga.mgh.harvard.edu/primerbank/index.html)
and Real Time PCR Primer Sets (http://www.realtimeprimers.org) for suitable, validated RT-PCR assays:)
Hi, I have a friend who desperately needs some nice ideas about a problem... He has some weird bacteria who are normally susceptible to Chloramphenicol, but he was able to isolate some...
09 October 2010 3,670 1 View
Your toughest technical questions will likely get answered within 48 hours on ResearchGate, the professional network for scientists.
07 August 2009 4,230 0 View
Hey, I wanted to test the RNA dependent RNA polymerase from NEB for a protocol I'm working on, it is a cool enzyme that produces dsRNA from ssRNA without need of any primer. The bad thing is,...
06 July 2009 9,330 0 View
Hi, I have been isolating some plasmids from E. coli for routine use (like PBBR1MCS), but I have a problem accurately quantifying the DNA that I get.. I used the Qiagen miniprep columns for 5...
06 July 2009 6,061 0 View
Hey, I am now trying to develop genetic systems for a bacterium, however it is pretty difficult to get DNA inside them somehow... I read the following article on Ultrasound mediated DNA (and...
05 June 2009 8,910 0 View
Dear i am work on RT-PCR, i have 2 bands on agarose gel (one at 150 BP and other at 400 BP), the 2 bands are parts from my gen after sequencing but now, i need to transfer all my product for form...
05 June 2009 10,010 0 View
Hi, I have to perform an experiment to see if my bacteria have strong restriction enzymes that are degrading the plasmids I am giving them... for that we had the idea to extract the bacterial...
04 May 2009 5,143 0 View
Would it be possible to use the Exonuclease I to remove unbound primers from a reverse-transcription reaction mix? I thought it might be suitable since cDNA and RNA are existing in a complex. Or...
03 April 2009 10,030 0 View
Hi, Does any of you have a good resource or protocol for PCR with a single primer? I took my protocol for double-primer RT-PCR but I think I am doing something wrong... And, supposedly the...
02 March 2009 3,353 7 View
Hi, Does anyone have an idea how to run ssDNA on agarose gels? I tried my regular protocol, but I am suspecting that the ssDNA is forming some secondary structures..
02 March 2009 505 1 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 View
i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...
05 August 2024 8,835 1 View
I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...
01 August 2024 2,079 0 View
Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...
31 July 2024 2,406 6 View
Can anyone with an MJ research / BioRad PCR machine from ~2010 or earlier tell me the external measurements (LxWxH) of the removable standard PCR alpha modules that can be removed from a PTC200 or...
30 July 2024 2,867 0 View
Hello all, I have been trying to follow a 2-stage PCR protocol used to amplify barcodes of a large yeast library, as per Nyugen et al. (2022) -...
30 July 2024 841 2 View
I need to install this program
25 July 2024 4,756 0 View
Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...
25 July 2024 4,927 3 View
I am preparing to run a lot of samples on a 384 well plate and I wanted to test if any well in my PCR machine is dead so as not to lose any data. I would really appreciate help with how I can do...
24 July 2024 8,172 1 View