Hello,
My name is Erica Sanchez. I have used your in silico PCR tool in the past
when designing primers for DNA amplification to check for single product
amplification. Recently, I have been designing RT PCR primers, across exon
boundaries to eliminate amplification of DNA contamination. I understand
that these primers should then NOT produce a product with the in silico PCR
tool when using the "genome assembly" as my target, but I realized that if I
changed the target to "UCSC genes" that the predicted product for amplifying
my cDNA does come up. I am emailing to ask for clarification of what UCSC
genes is referring to. Is this referring to RNA, cDNA, ESTs? I think it
makes sense that this is some king of cDNA library, but an unsure. All of my
RT primer pairs (designed across exon boundaries) are coming up with no
product under genome assembly and with my predicted amplification product
under UCSC genes target. Please provide me with clarification of this target
choice.
Thank you,
Erica Sanchez
*Example of Primers used:*
ELOVL 2 (gene name)
* F* 5'-CTCAGCTGGTGCAGTTCGTGC-3' * R* 5'- GGC TTT TTT CGG TAT GTC
TGA ACG-3'
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