RT-PCR

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. Lane 1 Lane 2,3 Lane 4,5 Lane 6,7 Lane 8,9 Lane 10,11 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.

Enhanced Avian HS RT-PCR kits Greater transcription lengths - up to 14 kb Detects low abundance mRNA Improved specificity and yield with JumpStart™ AccuTaq™ LA DNA polymerase Flexibility with One-Step & Two-Step Protocols HSRT20 HSRT100   Enhanced Avian First Strand Synthesis Kit Detects low abundance mRNA Optimum for transcribing through difficult secondary structure The resulting cDNA is ready for PCR amplification   Enhanced Avian Reverse Transcriptase Detects low abundance mRNA Best enzyme on the market for transcribing through difficult secondary structure More robust at elevated temperatures, up to 65°C AMV Reverse Transcriptase M-MLV Reverse Transcriptase JumpStart™ RED HT RT-PCR Kit

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