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 3,572 1 View
Your toughest technical questions will likely get answered within 48 hours on ResearchGate, the professional network for scientists.
07 August 2009 4,127 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 9,235 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 5,964 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 8,824 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 9,925 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 5,049 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 9,952 0 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 403 1 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 3,255 7 View
Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...
02 March 2021 4,029 3 View
I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...
01 March 2021 8,544 1 View
To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...
01 March 2021 8,683 4 View
Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?
01 March 2021 5,311 1 View
Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.
28 February 2021 5,008 4 View
I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...
27 February 2021 547 3 View
Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...
26 February 2021 4,777 4 View
Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...
26 February 2021 5,029 2 View
Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...
25 February 2021 6,969 3 View
After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...
25 February 2021 5,712 3 View