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.

Supplier:  Invitrogen™ 18080051

Catalog No. 18-080-051

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

Description

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 and the amount of starting material can vary from 1 pg to 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.

Using the SuperScript III First-Strand System
cDNA synthesis is performed in the first step using either total RNA or poly(A)⁺-selected RNA primed with oligo(dT), random primers, or a gene-specific primer. In the second step, PCR is performed in a separate tube using primers specific for the gene of interest. For the PCR reaction, we recommend one of the following DNA polymerases: Platinum Taq DNA Polymerase provides automatic hot-start conditions for increased specificity up to 4 kb, Platinum Taq DNA Polymerase High Fidelity provides increased yield and high fidelity for targets up to 15 kb, and Platinum Pfx DNA Polymerase provides maximum fidelity for targets up to 12 kb.

Order Info

Shipping Condition: Dry ice

Specifications

• Content And Storage:

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

• GC-Rich PCR Performance: High
• Technique: Reverse Transcription
• Reverse Transcriptase: SuperScript III
• For Use With (Application): Real Time PCR (qPCR), RT-PCR
• No. of Reactions: 50 Reactions
• Reagent Type: Reverse Transcription

• Format: Kit
• Reaction Speed: 50 min.
• Optimal Reaction Temperature: 50°C
• Shipping Condition: Dry Ice
• Final Product Type: First-Strand cDNA
• Reaction Format: Separate components
• Starting Material: RNA

For Research Use Only. Not for use in diagnostic procedures.