Invitrogen SuperScript III First-Strand Synthesis System
Optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA or RT-PCR
Manufacturer: Invitrogen 18080051
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
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Shipping Condition: Dry ice
|First-Strand cDNA Synthesis, Microarry analysis, RT-PCR|
|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. coliRNase 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|
|Store at -20deg.C|
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For Research Use Only. Not for use in diagnostic procedures.