The study of gene expression, at both the RNA and protein levels, often begins with the assay of low abundance transcripts and/or transcripts containing difficult secondary structure. These messages exceed the capabilities of most reverse transcriptases. Sigma's RT-PCR kits, based on the Enhanced Avian Reverse Transcriptase are the most effective tool for reproducible reverse transcription of low abundance messages and messages with difficult secondary structure, and for subsequent high fidelity amplification of the cDNA product.
Different poly (A)+ RNA were used as
a template in a two-step RT-PCR.
1 kb Ladder
2 kb Pol
3.5 kb Pol
5.3 kb TSC-2
6.8 kb Pol
8.9 kb APC
RT-PCR was performed on human phospholipase A2 (low abundance RNA, lane 2-7) and human HPRT RNA (a medium abundance RNA, lane 8-13) using Sigma's Enhanced Avian RT, RNase
H-reduced AMV RT and RNase
H-minus M-MLV RT.
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