Product description

TOPscript™ cDNA synthesis Kit (mix type/dN6 plus) is designed for the maximum-efficiency conversion of all types of RNA including mRNA to cDNA. The kit contains RNase inhibitor in TOPscript™ Reverse Transcriptase^R+ to protect RNA degradation and random hexamer (dN6) in 10X TOPscript™ RT reaction MIX (dN6). TOPscript™ Reverse Transcriptase is a highly thermostable version of M-MLV RT (RNase H-), and it can synthesize cDNA at 50~60℃ with highest fidelity. Thus, it is well suited to synthesize long cDNA (>20 kb) from long and highly secondary-structured RNA.

Characteristics

- cDNA synthesis across a broad temperature range

- Synthesis long cDNA

- Include all components required for cDNA synthesis

Applications

- First-strand of cDNA synthesis

- Primer extension

Supplied with

- TOPscript™ Reverse Transcriptase^R+ (200 U/μl)

- 10X TOPscript™ RT reaction MIX (dN6)

- 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.

Storage buffer

50 mM tris-HCl (pH 7.6), 150 mM NaCl, 1mM dTT, 0.1 mM EDTA, 0.1% NP-40, 50% glycerol

Standard reaction conditions

^a)Prepare one of the following RNA template.

    - Total RNA: 1 ng~5 μg

    - Messenger RNA (mRNA): 1 ng~250 ng

    - Specific RNA: 0.01 pg~0.5 μg

→ Incubate at 42℃~60℃ for 60 min.

→ Incubate at 95℃ for 5 min to inactivate the reaction.

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.

●  Enzynomics_EZ205_TOPscript™ cDNA synthesis Kit (mix type/dN6 plus)_manual

● Material Safety Data Sheet

- Enzynomics_MSDS_EZ205_TOPscript™ cDNA synthesis Kit (mix type/dN6 plus)_E