Invitrogen SuperScript First-Strand Synthesis System for RT-PCR
Optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA
Manufacturer: Invitrogen 11904018
The SuperScript First-Strand Synthesis System for RT-PCR can be used with as little as 1ng or as much as 5µg of total RNA. After synthesis, the cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations. In conjunction with PCR, the system can be used to detect the presence of rare messages, to quantitate the amount of specific mRNA from small numbers of cells, or to clone specific cDNAs without constructing an entire cDNA library. The system is flexible, allowing the use of any PCR enzyme. Combine with AccuPrime Taq DNA Polymerase or Platinum Taq DNA Polymerase for higher specificity PCR or with AccuPrime Pfx DNA Polymerase for high-fidelity cloning applications.
- SuperScript II RT The first-strand cDNA synthesis reaction is catalyzed by SuperScript II Reverse Transcriptase (RT). This enzyme has been engineered to reduce the RNase H activity that degrades mRNA during the first-strand reaction, resulting in greater full-length cDNA synthesis and higher yields of first-strand cDNA than obtained with RNase H+ RTs
- Because SuperScript II RT is not inhibited significantly by ribosomal and transfer RNA, it may be used effectively to synthesize first-strand cDNA from a total RNA preparation
- The enzyme exhibits increased thermal stability and may be used at temperatures up to 50°C
Using the SuperScript First-Strand Synthesis System for RT-PCR This system has been optimized to synthesize first-strand cDNA from varying amounts of starting material. The SuperScript II RT concentration has been lowered and RNaseOUT Recombinant RNase Inhibitor has been added to the system as part of this optimization process. Additionally, reaction conditions have been modified to further increase the sensitivity of the system. Using the kit, first-strand cDNA is synthesized using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer. PCR using primers specific for the gene of interest is performed in a separate tube.
PCR & Real-Time PCR, Reverse Transcription
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Shipping Condition: Wet ice
|First-Strand cDNA Synthesis, qRT-PCR|
|50μL Oligo(dT)12-18; (0.5μg/μL); 250μL Random hexamers (50ng/μL); 1mL 10X RT buffer; 500μL 25mM Magnesium Chloride; 250μL 0.1M DTT; 250μL 10mM dNTP mix; 50μL SuperScript II RT (50U/μL); 100μL RNaseOUT (40U/μL); 50μL E. coli RNase H (2U/μL); 1.2mL DEPC-treated water; 15μL Control RNA (50ng/μL); 20μL Control Primer A (10μM); 20μL Control Primer B (10μM)|
|Store at -20deg.C|
|Random Primers, Oligo dT Primers|
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For Research Use Only. Not for use in diagnostic procedures.