TOPscript™ Reverse Transcriptase

TOPscript™ Reverse Transcriptase is genetically engineered version of M-MLV RT which is highly thermostable, thus can synthesize cDNA at elevated temperatures up to 60°C. This property is very useful when RNA templates are long and have extensive secondary structures. TOPscript™ Reverse Transcriptase is capable of synthesizing cDNA longer than 20 kb from messenger RNA.

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TOPscript™ Reverse Transcriptase has wide optimal temperatures. Synthesis of cDNA from mRNA encoding glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was carried out by TOPscript™ Reverse Transcriptase (lanes 1-4) at 42, 50, and 60°C. The cDNA of GAPDH was also synthesized by other suppliers reverse transcriptases (lanes 5-16) for comparison purposes. Amplification of the GAPDH cDNA was performed by nTaq-Tenuto DNA Polymerase

Efficient synthesis of cDNA from long mRNA by TOPscript™ Reverse Transcriptase. Total RNA (100 ng) isolated from HeLa cells were first converted into cDNA using oligo (dT) by either TOPscript™ Reverse Transcriptase (lanes 1, 3, and 5) or Supplier A’s reverse transcriptase (lanes 2, 4, and 6). Reactions were carried out at 42°C, 50°C, and 60°C as indicated. Efficiency of LRP1 cDNA (14.9 kb) synthesis was verified by amplifying the 5’ end 0.9-kb region of the LRP1 cDNA using nTaq-Tenuto DNA Polymerase.