mRNA are easy to denature but denaturation of pre-mRNA requires additional steps.
I have tried heating at 70C and 80C for 10 minutes prior to cDNA synthesis, but it hasn't worked yet, can someone suggest another method?
Did you use RNAse H to degrade RNA from RNA:DNA hybride? It helped me.
I want to amplify a region of a specific pre mRNA, but the standard method is not working.
09 October 2013 9,704 6 View
I have tried isolating leukocytes from whole blood by Histopaque (sigma), but when I checked it in microscope, I found lots of platelets along with leukocytes, I tried washing, but it din't help much.
06 July 2013 1,950 1 View
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02 March 2012 7,453 0 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 try to make the Paal Knorr reaction between 2.5 hexnedione and 6-amino hexanoic acid, my problem is type of solvent to use, because 6-hexaminohexanoic acid is soluble in water but I am not...
02 March 2021 4,443 2 View
Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue
02 March 2021 5,433 5 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
Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...
01 March 2021 2,622 3 View
Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...
01 March 2021 8,169 2 View
I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...
28 February 2021 4,949 3 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