TOPscript™ cDNA Synthesis kit

Product description

TOPscript™ cDNA Synthesis Kit is designed for the highest efficiency conversion of RNA to cDNA. TOPscript™Reverse Transcriptase is genetically engineered version of M-MLV Reverse Transcriptase which is highly thermostable, thus can synthesize cDNA at elevated temperatures up to 60℃ A two-step RT-PCR format is useful for amplifying multiple targets from a single cDNA source, for maintaining archival cDNA, and for providing maximum flexibility in selecting a downstream qPCR reagent system. The TOPscript™ cDNA synthesis Kit is formatted for high-yield production of cDNA up to 20 kb and each reaction accommodates a range of RNA amounts from fg to μg.

Characteristics

- Robust cDNA synthesis across a broad range of temperatures from 42~60℃

- Produces excellent yields of amplifiable cDNA

- Can synthesize long cDNA targets, up to 20 kb, from small amounts of input RNA

- Able to reverse transcribe through RNA secondary structures

- Strict enzyme purity specifications

Applications

- cDNA cloning or cDNA library preparation

- Two step RT-PCR (end-point or real-time quantitative PCR)

Supplied with

- TOPscript™ Reverse Transcriptase (200 U/μl)

- 10X TOPscript™ RT Buffer

- RNase Inhibitor (40 U/μl)

- dNTP Mixture (2 mM)

- Oligo dT (80 μM)

- Random hexamer (100 μM)

- Sterile water (RNase free)

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- RNase-free

- Inhibitor-free

- Satisfactory yield and length of cDNA products

Unit definition

One unit is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble materrials using 0.4 mM poly(rA)-oligo(dT) as substrate at 37℃ in 10 min in a 1X reaction buffer. 

Note 

TOPscript™ Reverse Transcriptase performs optionally over the full range of 42℃~60℃. Typically, 50℃ is a good starting point. For RNAs containing secondary structure and other challenging targets, a synthesis temperature of 60℃ may be used without loss of performance.