ThermoScript™ RT-PCR System for First-Strand cDNA Synthesis

Catalog number: 11146024

Invitrogen™  Related applications: Reverse Transcription

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Description

The ThermoScript™ RT-PCR System for First-Strand cDNA Synthesis Kit is optimized for generating first-strand cDNA from total or poly(A)+ RNA. ThermoScript™ Reverse Transcriptase (RT) is an avian RT with reduced RNase H activity designed for RT-PCR of the most highly structured, difficult RNA templates. The ThermoScript™ RT-PCR systems are designed to copy RNA with secondary structure and to improve specificity of priming. In conjunction with PCR, the system can be used to detect the presence of rare messages or to quantitate the amount of specific mRNA from small numbers of cells. The systems contain ThermoScript™ RT, a cloned avian reverse transcriptase that has been engineered to have reduced RNase H activity and to have high thermal stability (up to 70°C). ThermoScript™ RT-PCR systems:

• Are optimized for RT-PCR
• Can be used with highly structured RNA or gene-specific primers
• Reduce RNase H activity for greater first-strand cDNA yield
• Amplify first-strand cDNA products directly without organic extractions or precipitations
• Include oligo(dT) and random hexamers to prime cDNA synthesis for flexible use

DNA polymerase options
The ThermoScript™ RT-PCR System for First-Strand cDNA Synthesis Kit offers a choice of Platinum® Taq DNA Polymerase for higher specificity when amplifying templates up to 3 kb, or Platinum® Taq DNA Polymerase High Fidelity for amplifying templates up to 12 kb.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Reverse Transcriptase: ThermoScript™
Downstream Application: Reverse Transcriptase PCR (RT-PCR)
Reaction Format: Separate Components
Optimal Reaction Temperature: 50-65°C
Primer Type: Oligo dT Primers, Random Primers
Product Size: 25 reactions
Shipping Condition: Dry Ice

Contents & storage

• 25 µl ThermoScript™ RT (15 U/µl)
• 500 µl 5X cDNA Synthesis Buffer
• 250 µl 0.1 M DTT
• 100 µl 10 mM dNTP Mix
• 25 µl RNaseOUT™ (40 U/µl)
• 25 µl Oligo (dT)20 (50 µM)
• 50 µl Random Hexamers (50 ng/µl)
• 1.25 ml DEPC-Treated Water
• 50 µl E. coli RNase H (2 U/µl)

Store at -20°C (stability can be extended by storage at -70°C).

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Manuals & protocols

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