Cambio - Excellence in Molecular Biology

In vitro Transcription

In vitro Transcription: In Vitro Transcription

MonsterScript™ 1st-Strand cDNA Synthesis Kit

MonsterScript 1st-Strand cDNA Synthesis Kit is optimized for generating full-length first-strand cDNA from mRNA contained in a total cellular RNA sample or from purified Poly(A) RNA

Epicentre® Biotechnologies

MonsterScript™ 1st-Strand cDNA Synthesis Kit

MonsterScript 1st-Strand cDNA Synthesis Kit is optimized for generating full-length first-strand cDNA from mRNA contained in a total cellular RNA sample or from purified Poly(A) RNA

Epicentre® Biotechnologies

The MonsterScript™ 1st-Strand cDNA Synthesis Kit is optimized for generating full-length first-strand cDNA from multiple mRNA species in samples containing total cellular or purified polyadenylated RNA. 

MonsterScript™ 1st-Strand cDNA Synthesis Kit uses MonsterScript™ Reverse Transcriptase, a reverse transcriptase that lacks RNase H activity. The enzyme's lack of RNase H activity contributes to its ability to make longer cDNAs and more complete libraries of first-strand cDNA molecules. EPICENTRE scientists have demonstrated reverse transcription of 15-kb RNA templates using MonsterScript Reverse Transcriptase. 

MonsterScript Reverse Transcriptase produces full-length cDNA from mRNA up to or greater than 15kb MonsterScript™ Reverse Transcriptase produces full-length cDNA from mRNA up to or greater than 15 kb The ≈15.2-kb human guanine nucleotide exchange factor, p532, mRNA ('HGNEFp532 mRNA') was reverse transcribed using total HeLa cellular RNA and an oligo(dT)-containing primer in a standard MonsterScript reaction. Two µl of the reverse transcription reaction was used to amplify a 1.3-kb region within 68 bases of the 5'-end of the HGNEFp532 mRNA with the FailSafe™ PCR System. 1A. Detection of the PCR product demonstrates full-length reverse transcription of the HGNEFp532 mRNA by MonsterScript Reverse Transcriptase. 1B. Agarose gel analysis of the PCR reaction shows the 1.3-kb amplicon amplified from the extreme 5'-end of the HGNEFp532 mRNA.
MonsterScript Reverse Transcriptase produces full-length cDNA from mRNA up to or greater than 15kb

The kit also contains MonsterScript™ RT PreMix, which contains optimized concentrations of dNTPS, Mg 2 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 a random nonamer primer.

Figure 2. MonsterScript Reverse Transcriptase was used to reverse transcribe six dilutions Figure 2. MonsterScript™ Reverse Transcriptase was used to reverse transcribe six dilutions (8 x 10-6 to 8 x 10-1pg) 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 PCR reactions, in duplicate, using TAQurate™ GREEN Real-Time PCR MasterMix. A PCR efficiency of 99.4% and correlation coefficient of 1.000 was achieved.

*Covered by issued and/or pending patents.




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MonsterScript™ 1st-Strand cDNA Synthesis Kit

MonsterScript 1st-Strand cDNA Synthesis Kit is optimized for generating full-length first-strand cDNA from mRNA contained in a total cellular RNA sample or from purified Poly(A) RNA

Epicentre® Biotechnologies

Protocols for: MonsterScript™ 1st-Strand cDNA Synthesis Kit & MonsterScript™ Reverse Transcriptase 

Due to the constant updating of the protocols by the manufacturer we have provided a direct link to Epicentre’s product page, where the latest protocol is available.

Please note this will open a new page or window on your computer.

 

MonsterScript™ 1st-Strand cDNA Synthesis Kit Protocol

(catalogue number MS040910 / MS041050) 

MonsterScript™ Reverse Transcriptase 

(catalogue number MSTA5124 / MSTA5110) 

Please note: all protocols off site are the responsibility of the products supplier

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

MonsterScript™ 1st-Strand cDNA Synthesis Kit

MonsterScript 1st-Strand cDNA Synthesis Kit is optimized for generating full-length first-strand cDNA from mRNA contained in a total cellular RNA sample or from purified Poly(A) RNA

Epicentre® Biotechnologies

References:

    1. Vaidyanathan, R. (2005)
    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.
  • Chen, W., et al. (2010) Disruption of ptLPD1 or ptLPD2, Genes That Encode Isoforms of the Plastidial Lipoamide Dehydrogenase, Confers Arsenate Hypersensitivity in Arabidopsis, Plant Physiology 153 , 1385-1397.
  • Hernandez-Diaz, I., et al. (2010) The Mineralocorticoid Receptor Is a Constitutive Nuclear Factor in Cardiomyocytes due to Hyperactive Nuclear Localization Signals, Endocrinology 151 , 3888-3899.
  • Teterina, N. L., et al. (2010) Viable Polioviruses That Encode 2A Proteins with Fluorescent Protein Tags, J. Virol. 84 , 1477-1488.
  • Friedman, S. D., et al. (2009) Gene Mapping and Phylogenetic Analysis of the Complete Genome from 30 Single-Stranded RNA Male-Specific Coliphages (Family Leviviridae), J. Virol. 83 , 11233-11243.
  • Kent, T., et al. (2009) The 5' external transcribed spacer in mouse ribosomal RNA contains two cleavage sites, RNA 15 , 14-20.
  • Budde, P. P., et al. (2007) Characterization of a higBA Toxin-Antitoxin Locus in Vibrio cholerae, J. Bacteriol. 189 , 491-500.
  • Chumley, M. J., et al. (2007) EphB Receptors Regulate Stem/Progenitor Cell Proliferation, Migration, and Polarity during Hippocampal Neurogenesis, J. Neurosci. 27 , 13481-13490.
  • Alice, A. F., et al. (2006) Genetic and Transcriptional Analysis of the Siderophore Malleobactin Biosynthesis and Transport Genes in the Human Pathogen Burkholderia pseudomallei K96243, J. Bacteriol. 188 , 1551-1566.

 

EPICENTRE Forum 12 (1)Alice, A.F. et al. (2006) J. Bacteriol. 188, 1551.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

MonsterScript™ 1st-Strand cDNA Synthesis Kit

MonsterScript 1st-Strand cDNA Synthesis Kit is optimized for generating full-length first-strand cDNA from mRNA contained in a total cellular RNA sample or from purified Poly(A) RNA

Epicentre® Biotechnologies

Applications 

  • First-strand cDNA synthesis
  • Production of cDNA 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

 

Benefits 

  • MonsterScript™ Reverse Transcriptase lacks RNase H, enabling improved synthesis of full-length cDNA even for long mRNA
  • MonsterScript is 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, Mg 2 and Betaine for superior performance and minimal pipetting steps
  • Betaine in the cDNA Synthesis PreMix reduces pausing and stops during reverse transcription
  • The kit includes both an oligo(dT)-containing and a random nonamer primer
  • First-strand cDNA can be made from picogram amounts of total RNA
  • A potent RNase Inhibitor is included

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200