Invitrogen™ SuperScript™ III First-Strand Synthesis System

Optimized to synthesize first-strand cDNA from purified poly(A)⁺ or total RNA or RT-PCR

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

RNA targets from 100 bp to >12kb can be detected with this system. The amount of starting material can vary from 1pg to 5μg 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° to 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, one of the following DNA polymerases is recommended
• Platinum Taq DNA Polymerase provides automatic hot-start conditions for increased specificity up to 4kb
• Platinum Taq DNA Polymerase High Fidelity provides increased yield and high fidelity for targets up to 15kb
• Platinum Pfx DNA Polymerase provides maximum fidelity for targets up to 12kb

• Optimal reaction temperature: 50°C
• Sufficient for 50 reactions, based on a 20μL reaction size

PCR & Real-Time PCR, Reverse Transcription

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.
  
  

• Format: First-Strand Synthesis System
• No. of Reactions: 50 Reactions
• Reaction Format: Separate components
• Reagent Type: Reverse Transcription
• Technique: Reverse Transcription
• Optimal Reaction Temperature: 50°C
• Shipping Condition: Dry Ice

• Final Product Type: First-Strand cDNA
• GC-Rich PCR Performance: High
• PCR Method: RT-PCR, RT-qPCR
• Reaction Speed: 50 min.
• Starting Material: RNA
• Volume: 20 μL
• Reverse Transcriptase: SuperScript III
• For Use With (Application): Microarray Analysis, Reverse Transcriptase PCR (RT-PCR)

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