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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

Encompass

Catalog No. 18-080-051


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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

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

Specifications

• 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.

First-Strand Synthesis System
50 Reactions
Separate components
Reverse Transcription
Reverse Transcription
50°C
Dry Ice
First-Strand cDNA
High
RT-PCR, RT-qPCR
50 min.
RNA
20 μL
SuperScript III
Microarray Analysis, Reverse Transcriptase PCR (RT-PCR)
SDS
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