This product will be discontinued on September 1, 2016. For superior performance, we recommend you switch to the SuperScript® IV First-Strand Synthesis System and Platinum® Taq DNA Polymerase High Fidelity.

ThermoScript™ RT-PCR System & Platinum® Taq DNA Polymerase High Fidelity

Catalog number: 11146040

Invitrogen™  Related applications: Reverse Transcription

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Description

The ThermoScript™ RT-PCR System is designed for the detection and analysis of RNA in a two-step process. ThermoScript™ reverse transcriptase (RT) is used to generate full-length cDNA transcripts using either total RNA or poly(A)+ RNA. PCR amplification is then performed using Platinum® Taq DNA Polymerase High Fidelity.
cDNA Yield—Low RNase H activity of ThermoScript™ RT maximizes first-strand cDNA yield
Thermostability—ThermoScript™ RT is active at temperatures up to 70°C
Fidelity—Platinum® Taq DNA Polymerase High Fidelity offers 6 times the fidelity of Taq DNA Polymerase alone
Amplicon Size—Platinum® Taq DNA Polymerase High Fidelity is ideal for templates from 100 bp to 15 kb (Figure 1)

High-Yield First Strand Synthesis
ThermoScript™ Reverse Transcriptase (RT) is an avian RT with reduced RNase H activity designed for RT-PCR of your most highly structured, difficult RNA templates. ThermoScript™ RT-PCR Systems are designed to read through secondary structures and to improve priming specificity. Priming may be performed using oligo(dT), or random primers (both included), or a gene-specific primer. In conjunction with PCR, the system can be used to detect the presence of rare messages, or to quantitate the amount of specific mRNAs from small numbers of cells.

High-Fidelity PCR Amplification
Platinum® Taq DNA Polymerase High Fidelity is ideal for the amplification of DNA fragments, when high yields and robust amplification are required. Fidelity is maximized through the use of a mixture of Platinum® Taq DNA Polymerase and the proofreading (3’-5’ exonuclease activity) enzyme Pyrococcus species GB-D polymerase. PCR specificity is improved with the incorporation of Platinum® automatic "hot-start" technology.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

PCR Method: 2-Step RT-PCR
Reverse Transcriptase: ThermoScript™
Polymerase: Platinum® Taq DNA Polymerase High Fidelity
Hot Start: Built-In Hot Start
Sample Type (General): RNA
Product Size: 100 reactions
Detection Method: Gel Electrophoresis
Final Product Size(s): 12 kb or less
Primer-Probe Compatibility: Unlabeled Oligos

Contents & storage

• 100 units Platinum® Taq DNA Polymerase High Fidelity
• 1.0 ml 10X High Fidelity PCR Buffer
• 1.0 ml 50 mM Magnesium Sulfate
• 100 µl ThermoScript™ RT (15 U/µl)
• 500 µl 5X cDNA Synthesis Buffer
• 250 µl 0.1 M DTT
• 2 × 250 µl 10 mM dNTP Mix
• 100 µl RNaseOUT™ (40 U/µl)
• 100 µl Oligo (dT)20 (50 µM)
• 250 µl Random Hexamers (50 ng/µl)
• 1.25 ml DEPC-Treated Water
• 2 × 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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