SuperScript® III First-Strand Synthesis SuperMix

Catalog number: 18080400

Invitrogen™  Related applications: Reverse Transcription

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The SuperScript® III First-Strand Synthesis SuperMix is an optimized SuperMix formulation for first-strand cDNA synthesis 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 0.1 pg to 5 µg of total RNA.

Using the SuperScript® III First-Strand Synthesis SuperMix
The kit includes SuperScript® III/RNaseOUT™ Enzyme Mix, 2X First-Strand Reaction Mix, and Annealing Buffer. SuperScript® III Reverse Transcriptase, included in the Enzyme Mix, is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can be used to synthesize cDNA at a temperature range of 45–55°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 can be used to synthesize cDNA from total RNA. RNaseOUT™ Recombinant RNase Inhibitor is included in the Enzyme Mix to safeguard against degradation of target RNA due to ribonuclease contamination. The 2X First-Strand Reaction Mix includes 10 mM MgCl2 and 1 mM of each dNTP in a buffer formulation that has been optimized for first-strand synthesis of cDNA. The Annealing Buffer is used in the initial template-primer annealing step. Separate tubes of oligo(dT)20 and random hexamers are also provided. cDNA synthesis can be performed using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer.
For Research Use Only. Not for use in diagnostic procedures.


Reverse Transcriptase: SuperScript® III
Downstream Application: Reverse Transcriptase PCR (RT-PCR), Real-Time Quantitative PCR (qPCR)
Reaction Format: SuperMix or Master Mix
Primer Type: Oligo dT Primers, Random Primers
Product Size: 50 reactions
Shipping Condition: Dry Ice
Optimal Reaction Temperature: 50° C

Contents & storage

• 2X Reaction Mix (500 µl)
• SuperScript® III Enzyme Mix (100 µl)
• Oligo(dT)20 (50 µl total provided at 50 µM)
• Random hexamers (50 µl total provided at 50 ng/µl)
• Annealing buffer (50 µl)

Enough material provided for 50 reactions (20 µl each). Store at -20°C.


Manuals & protocols

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