Invitrogen™ SuperScript™ III First-Strand Synthesis System

Invitrogen™ SuperScript™ III First-Strand Synthesis System

The SuperScript™ III First-Strand Synthesis System for RT-PCR is optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA. RNA targets from 100 bp to >12 kb can be detected with this system. The amount of starting material can vary from 1 pg–5 µg of total RNA. SuperScript™ III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme is used to synthesize cDNA at a temperature range of 42–55°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript™ III RT is not significantly inhibited by ribosomal and transfer RNA, it may be used to synthesize first-strand cDNA from a total RNA preparation.

Contents

• Oligo(dT)20 (50 µM), 50 µl
• Random hexamers (50 ng/µl), 250 µl
• 10X RT buffer, 1 mL
• 0.1 M DTT, 250 µl
• 25 mM magnesium chloride, 500 µl
• 10 mM dNTP mix, 250 µl
• SuperScript III RT (200 U/µl), 50 µl
• RNaseOUT (40 U/µl), 100 µl
• E. coli RNase H (2 U/µl), 50 µl
• DEPC-treated water, 1.2 mL
• Total HeLa RNA (10 ng/µl), 20 µl
• Sense Control Primer (10 µM), 25 µl
• Antisense Control Primer (10 µM), 25 µl