KR107-TIANScript II RT Kit

TIANScript II RT Kit
——For complex secondary structures and
long first-strand cDNA synthesis

TIANScript II RT Kit
Cat.No. 4992910/4992911

Storage
All the components can be stored at -30~-15°C for up to 12 months.

Introduction
TIANScript II RT Kit supplies all the required reagents for first-strand cDNA
synthesis, including the TIANScript II RTase which is modified from M-MLV,
with higher affinity to template and lower RNase H activity. Therefore
it has the ability to read complex secondary structure of template and
reverse transcribe long RNA in the process of first-strand cDNA synthesis.
The 5×TIANScript II RTase Buffer in the kit not only guarantees high
enzyme activity of new RTase, but also expands the template input range.
cDNA synthesized by this kit has high quality and is good for downstream
analysis.

Product Highlights
High enzyme activity: High RTase activity, which is compatible for
downstream analysis.
Wide range of substrates:Suitable for reverse transcription of all kinds of
RNA, especially for RNA which has complex secondary structure.
Long RT fragments: The maximum length of first-strand cDNA the kit can
synthesize is up to 12 kb.

Applications

1. Synthesis of first-strand cDNA.
2. Construction of cDNA library.
3. Two step RT-PCR.
4. RACE analysis.

Source of Enzyme
Recombinant E. coliwhich contains improved M-MLV gene.

Unit Definition

1. 1 unit (U) is defined as the amount of enzyme required to catalyze the
   incorporation of 1 nM of dNTP into acid-insoluble in 10 min at 37°C,
   using polyA.poly(dT)12-18 as template and primers.

Important Notes Before Starting

1. Solutions that used for cDNA synthesis should be treated with DEPC
   and autoclaved before use. For reagents not suitable for autoclaving,
   prepare the reagents with the sterilized container and water, and then
   filter to obtain the final solutions.
2. Avoid genomic DNA contamination in RNA sample.
3. Repeated freezing and thawing of RNA should be avoided. Keep the
   dissolved RNA on ice.
4. All the components of the kit should be stored at -30~-15°C.
5. When using Random Primers, please note the ratio of the amount
   of primers and the amount of RNA. It is suggested to use 50 ng of
   Random Primers for every 5 µg total RNA template. Increase the ratio of
   Random/RNA will improve the synthesis of short cDNA (~500 bp); and
   decrease the ratio of Random/RNA will improve the synthesis of long
   cDNA.
6. For RNA templates rich in secondary structures, it is suggested to heat
   at 65°C for 5 min before the reaction.
7. If the reverse primer of PCR reaction would be used in reverse
   transcription, in order to avoid nonspecific amplification led by primer
   mismatch, the whole transcription process could be done at 50°C.
8. We suggest to terminate the synthesis at 70°C for 15 min in order to
   avoid cDNA damage.
9. When using the enzymes, please mix enzymes gently to avoid bubbles;
   please centrifuge carefully to collect the enzyme before pipetting, and
   please aspirate slowly since enzymes are sticky.
   Note: For long cDNA synthesis, please prepare fresh, integrate and
   pure RNA template.

KR107-TIANScript II RT Kit

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