Ammon Thompson

3 Questions 12 Answers 0 Followers

Questions related from Ammon Thompson

What is the most accurate method of comparing a gene's expression level between tissue types using Real Time qPCR?

I need to see if a gene has higher expression in one tissue type compared to another. I would prefer not to use housekeeping genes for normalization.

09 September 2012 1,638 27 View

For reverse transcription, is there a difference between using oligo (dT)12-18 and oligo(dT)20? Does it matter for real time quantitative PCR?

All the different sizes of oligo dT seem to have really low Tm (32-39 C) and I've been running my RT reactions at 50 degrees which is around 15 degrees above their Tm values. Is the length of the...

04 April 2012 7,127 5 View

TaqMan no-RT controls showing amplification that is not from genomic DNA in real time quantitative PCR.

I ran the negative control RT reaction (no RT enzyme) using random hexamers and oligo dT primers. I then used that same sample for qPCR of three different genes in fish skeletal muscle. What is...

03 March 2012 3,502 16 View