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MonsterScript™ Reverse Transcriptase

MonsterScript™ Reverse Transcriptase

货号

MSTA5110

规格

10 Rxns

存储温度

-20℃

产品详情

Applications

  • First-strand cDNA synthesis for subsequent PCR or real-time PCR.
  • RT-PCR validation of gene expression data obtained from microarray experiments.
  • RT-PCR validation and quantification of gene silencing by RNA interference.
  • Preparation of labeled cDNA probes.

MonsterScript™ Reverse Transcriptase is a thermostable reverse transcriptase that completely lacks RNase H activity. The enzyme's lack of RNase H activity contributes to its ability to make longer cDNAs and more complete full-length libraries of first-strand cDNA molecules compared to other reverse transcriptases. EPICENTRE scientists have demonstrated reverse transcription of 15-kb RNA templates using MonsterScript™ Reverse Transcriptase (Fig. 1).

The MonsterScript 1st-Strand cDNA Synthesis Kit includes MonsterScript RT PreMix, which contains optimized concentrations of dNTPs, Mg2+, and betaine.* Betaine reduces pausing and stops by the reverse transcriptase, enabling improved reverse transcription through difficult sequences, such as regions of high GC content. The kit also contains both an oligo(dT)-containing primer and random nonamer primers.

Benefits

  • No RNase H activity, enabling improved synthesis of full-length cDNA even for long mRNA, using random priming.
  • Thermostable, permitting reverse transcription at temperatures >50°C, which reduces RNA secondary structure and improves priming specificity.
  • MonsterScript RT PreMix contains optimized concentrations of dNTPs, Mg2+, and betaine for superior performance and minimal pipetting steps.
  • The kit includes both an oligo(dT) and random nonamer primers for maximal transcript coverage.
  • First-strand cDNA can be made from picogram amounts of total RNA (Fig. 2).
  • The kit includes a potent RNase Inhibitor to protect the integrity of template RNA.

 

Figure 1

   Figure 2
Figure 1. MonsterScript™ Reverse Transcriptase produces full-length cDNA from mRNA greater than 15 kb. The ≈15.2-kb HGNEFp532 mRNA was reverse-transcribed from total HeLa RNA in a standard MonsterScript reaction. Two microliters of the reaction was used to PCR amplify a 1.3-kb region within 68 bases of the 5´ end of the HGNEFp532 mRNA (A). Agarose gel electrophoresis of the 1.3-kb amplicon from the 5´ end of the mRNA demonstrates full-length cDNA synthesis (B).   Figure 2. MonsterScript™ Reverse Transcriptase was used to reverse-transcribe six dilutions (8 x 10-6 to 8 x 10-1 pg) of an RNA transcript into cDNA in 20-µl reactions. Ten microliters of each reverse transcription reaction was then used in 50-µl real-time PCRs, in duplicate, using EPICENTRE's TAQurate™ GREEN Real-Time PCR MasterMix. A PCR efficiency of 99.4% and correlation coefficient of 1.000 was achieved.

Product Citations

  1. Alice, A.F. et al. (2006) J. Bacteriol. 188, 1551.
  2. Budde, P.P. et al. (2007) J. Bacteriol. 189, 491.
  3. Conejo, M. et al. (2008) J. Molec. Evolution 66, 11.
  4. Chumely, M.J. et al. (2007) J. Neurosci. 27, 13481.
  5. Borchert, G.M. (2006) Nature Struct. Mol. Biol. 13, 1097.

*Covered by issued and/or pending patents.

 

Catalog No. Size
MonsterScript™ 1st-Strand cDNA Synthesis Kit
MS040910 10 Reactions
MS041050 50 Reactions
Contents: MonsterScript™ Reverse Transcriptase (50 U/µl) with RNase Inhibitor, MonsterScript™ 5X cDNA PreMix (contains buffer, dNTPs, and betaine*), V3-Oligo(dT)21 Primer (10 µM), Random 9-mer Primers, RNase-Free Water.
MonsterScript™ Reverse Transcriptase
MSTA5110 10 Reactions
MSTA5124 24 Reactions
Includes MonsterScript™ 5X Reaction Buffer.

 

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