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ExactSTART™ End-Tagged Double-Strand cDNA Synthesis Kit for Small RNA

货号

ES81010

规格

1Kit

存储温度

-70℃

产品详情

Applications

Generation of dsDNA from small RNAs for:

High-throughput (next-gen) sequencing templates.
Small RNA RT-PCR templates.
Labeled target for small RNA microarray analyses.
Small RNA libraries for cloning.

The ExactSTART™ End-Tagged Double-Strand cDNA Synthesis Kit for Small RNA* selectively tags and converts small RNAs that contain 5´-monophosphate and 3´-hydroxyl ends (5´ pN—OH 3´), or 5´-triphosphate and 3´-hydroxyl ends (5´ pppN—OH 3´) to amplified dsDNA. Species of RNA tagged by the kit include mammalian miRNA, other small noncoding RNAs, and small primary transcripts.

An optimized RNA ligation reaction is used to tag the 5´ end, and a high-efficiency polyadenylation/cDNA synthesis reaction is used to tag the 3´ end of the RNAs (Fig. 1). Intervening size-separation by gel electrophoresis is not required. An optional step, using a unique RNA 5´ Polyphosphatase* enzyme (discovered and characterized by EPICENTRE scientists) enables tagging and dsDNA synthesis of small primary transcripts with 5´-triphosphorylated ends. An initial, simple RNA size-selection procedure removes large RNAs such as mRNA, and 18S and 28S rRNA, to ensure that only small RNAs are converted to dsDNA.

Benefits

Strong selection process yields libraries with a very high proportion of dsDNA derived from small (<200-base) RNAs with 5´-monophosphorylated or 5´-triphosphorylated ends.
Rapid and simple RNA size-selection procedure without the need for columns or gel electrophoresis.
One-day protocol.
User-defined PCR primers can be used to generate dsDNA for a specific downstream application (Fig. 1).

Figure 1 (click to enlarge). Schematic overview of the process for the ExactSTART™ End-Tagged Double-Strand cDNA Synthesis Kit for Small RNA.

*Patent pending

Catalog No.	Size

ExactSTART™ End-Tagged Double-Strand cDNA Synthesis Kit for Small RNA
ES81010	10 Reactions
Contents: RNA 5´ Polyphosphatase and Buffer, Poly(A) Polymerase and Buffer, Glycogen, RNase Inhibitor, ATP, T4 RNA Ligase and Buffer, 10X Ligase and RT Buffer, Small RNA Acceptor Oligo, RNase Solution, MMLV-RT, Small RNA cDNA Synthesis Primer, Small RNA PCR Primer 1, Small RNA PCR Primer 2, FailSafe™ 2X PreMix E, Small RNA Enrichment Solution, Nuclease-Free Water, DTT, Control RNA Template.

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