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11117831001

Roche

cDNA Synthesis Kit

kit of for up to 10 reactions

Synonym(s):
RT-PCR

Quality Level

manufacturer/tradename

Roche

packaging

kit of for up to 10 reactions

General description

Optimized one-tube procedure for the synthesis of double-stranded cDNA up to 3 kb from total RNA or mRNA.
The cDNA Synthesis System uses an optimized one-tube procedure, based on the method of Gubler and Hoffmann.
Sample: 1 - 20 μg total RNA, 0.3 - 2 μg mRNA
Assay time: 60 minutes for 1st strand synthesis, 120 minutes for 2nd strand synthesis
Product: double-stranded cDNA copy of an RNA sample

Application

Optimized one-tube procedure for the synthesis of double-stranded cDNA from total RNA or mRNA.
The ds cDNA can be used for the construction of non-directional cDNA libraries, as a starting point for substractive hybridization experiments to enrich differentially expressed genes, and for the in vitro transcription of whole cDNA populations to generate labeled cRNA for hybridization on DNA microarrays.

Components

  • AMV Reverse Transcriptase Buffer, 5x concentrated
  • AMV Reverse Transcriptase (25 U/μl)
  • 1,4-Dithiothreithol (0.1 M)
  • Protector RNase Inhibitor (25 U/μl)
  • Oligo (dT)15 primer, 200 μM (1 μg/μl)
  • Oligo [(dT24 )T7promotor]65 primer, 100 μM (2 μg/μl)
  • dNTP mixture (each nucleotide, 10 mM)
  • Control neo mRNA (0.2 μg/μl)
  • 2nd strand synthesis buffer, 5x concentrated
  • 2nd strand enzyme blend (mixture of DNA Polymerase I, E. coli Ligase and RNase H)
  • T4 DNA Polymerase (1 U/μl)
  • Double-distilled water
  • RNase I (10 U/μl)
  • Proteinase K, recombinant, PCR grade (50 U/mll)

Other Notes

For life science research only. Not for use in diagnostic procedures.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

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Certificate of Origin

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