Truseq mRNA PCR cycles

[Seq-MDC]

Dear all

i was wondering if anybody can help answering this Question?
I do estimate the number of the PCR Cycles should be used to amplify the Ligated Product according to the qPCR Quantification using these methode: take 1 μl of you adapter-*‐ligated DNA and mix it with 1 μl of TruSeq PCR Quantification Primer Mix (25 μM), 8 μl H2O and 10 μl 2X SYBR Green Master Mix. Run on qPCR machine with same perimeters as used for library amplification with 25 cycles.
Determine the number of cycles it took to get to 50% of amplication (late log phase)and use that number of cycles minus minus X.
I found out same times that the number of the cycles needed for some of the samples is 6 cycles!
i m not sure which is the minimum of the number of cycles you should not go under? any experiences about this issue? i will be appreciate if i get some feedback. Thank you

[ZWB]

While 6 cycles does seem really low, I've done as few as 5 cycles with good results. If you are worried you could do an extra cycle or two (no more that 8 total) and the library should still be OK. I've found the best thing is to trust the qPCR, if it indicates 6 cycles is optimal I would use that number.

[Seq-MDC]

Thanks ZWB
i think too that we can trust the q-PCR