t**s
发帖数: 284 | 1 I think different softwares score primers differently. But i don't think
they can be too much off each other.
Primers are dirt cheap. Why don't you design 2 pairs out of a software
and do an annealing temperature optimization? Or maybe run touch-down PCR.
RE map is restriction enyme map. You cut your PCR products with "RE"
and run on a gel to see if the cut produces correct DNA fragment bands. | f*****a
发帖数: 7 | 2 I only use SYBR. What I did is
1. Use Primer Express 1.5 or 2.0 from ABI to design primers. Follow their
guidelines. Or just use Primer3.
2. Use Amply 1.2 (or higher) and Netprimer http://www.premierbiosoft.com/n
etprimer/) to evaluate candidates.
3. Blast your primers.
4. Pick up 2-3 pairs. Do standard and dissociation curves. Chose the one wor
k best.
Make sure to chose the right normalization primers for your experiment. 18S,
actin, GAPDH, tubulin... |
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