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Tetro™ cDNA Synthesis Kit contains all the necessary components to generate cDNA from an RNA template. The generated cDNA is suitable for PCR with gene-specific primers or for other downstream applications. The kit contains MMLV Reverse Transcriptase and is suitable for first strand cDNA synthesis, cDNA library construction and the production of templates for PCR amplification.
Tetro™ cDNA Synthesis Kit contains our highly sensitive MMLV reverse transcriptase, oligo (dT)18 and random hexamer primers and all the necessary components to generate high quality cDNA from RNA templates (fig. 1). The first-strand cDNA generated is ideal for PCR (fig. 2) and can be used in a variety of other applications, such as analyses of cellular RNAs, characterization of RNA splice variants and the generation and cloning of cDNA.
Tetro cDNA Synthesis Kit is optimized for RT reactions over a wide range of total RNA concentrations (10 pg-5 μg), such that long and low-abundance cDNAs can be detected by amplification after cDNA synthesis. The kit contains oligo (dT)18 and random hexamer primers. The kit components are fully optimized to generate maximum yields of full-length cDNA.
Fig. 1 High sensitivity on mouse total RNA.
A ten-fold serial dilution of RNA (1 μg to 100 pg) was reverse transcribed using Tetro Reverse Transcriptase and oligo (dT)18. The resultant cDNA was then used as template in a PCR using primers for amplification of a 700 bp fragment from mouse b-actin. Lanes 1-5 correspond to PCR product from the serial dilution above, reactions were carried out in duplicate. HyperLadder 50bp (M).
Fig. 2 High sensitivity on human DNA.
A ten-fold serial dilution of RNA (1 μg to 10 pg) was reverse transcribed using Tetro Reverse Transcriptase and oligo (dT)18. The resultant cDNA was then used as a template in a PCR using primers for amplification of a 470 bp fragment from human GAPDH. Lanes 1-5 correspond to PCR product from the serial dilution above, reactions were carried out in duplicate. HyperLadder 50bp (M).
Reagent |
30 reactions |
100 reactions |
5x RT Buffer |
120 µL |
1.2 mL |
Reverse Transcriptase (200 u/µL) |
30 µL |
100 µL |
RNase Inhibitor (10 u/µL) |
30 µL |
100 µL |
dNTP Mix 10 mM Total |
30 µL |
100 µL |
Oligo (dT)18 Primer Mix |
30 µL |
100 µL |
Random Hexamer Primer Mix |
30 µL |
100 µL |
DEPC-treated Water |
1.8 mL |
1.8 mL |
All kit components should be stored at -20°C upon receipt. When stored under the recommended conditions and handled correctly, full activity of the kit is retained until the expiry date on the outer box label.
Tetro cDNA Synthesis Kit is shipped on dry/blue ice