Hi all,
I used same cdna template.. but when i optimize my primer using conventional pcr method, i did annealing temp adjustment and mgcl optimization. the problem is sometime i got pretty nice bands, but sometimes i got poor amplification or even no amplification, even i used same primer stock, same mgcl2 and everything is the same. it's really odd how could this occur. any advise? thanks
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susanna
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Posted 05 March 2010 - 01:08 AM
Poor cDNA quality, i would say
wntiong, on Mar 2 2010, 03:51 AM, said:
Hi all,
I used same cdna template.. but when i optimize my primer using conventional pcr method, i did annealing temp adjustment and mgcl optimization. the problem is sometime i got pretty nice bands, but sometimes i got poor amplification or even no amplification, even i used same primer stock, same mgcl2 and everything is the same. it's really odd how could this occur. any advise? thanks
I used same cdna template.. but when i optimize my primer using conventional pcr method, i did annealing temp adjustment and mgcl optimization. the problem is sometime i got pretty nice bands, but sometimes i got poor amplification or even no amplification, even i used same primer stock, same mgcl2 and everything is the same. it's really odd how could this occur. any advise? thanks
