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文献和实验PCR引物设计和优化(PCR PRIMER DESIGN AND REACTION OPTIMISATION)
is supposed to work well in reverse transcriptase buffer, and vice-versa, meaning 1-tube protocols (with cDNA synthesis and subsequent PCR) are possible (Krawetz et al., 19xx; Fuqua et al., 1990)。 Higher than 50mM KCl or NaCl inhibits Taq
Denature the RNA at 650 C, 5min., then ice. Dilute Oligo-dTplus (T12NA, T12NT, T12NG, T12NC) primers to final concentration 50uM. Reverse transcription (in 4 tubes): 10ul RNA Water 1ul DNA-free total RNA
, T12NC) primers to final concentration 50uM. Reverse transcription (in 4 tubes): 10ul RNA Water 1ul DNA-free total RNA 4ul 5Xreverse transcription buffer 2ul 0.1M DTT 1ul 10mM dNTP stock
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