cDNA Synthesis Kit

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Catalogue No: abx460012
Price: US$304.50
(Size: 50 rxns)
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cDNA Synthesis Kit

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Datasheet
cDNA Synthesis Kit for optimal cDNA synthesis from a variety of RNA samples over a wide temperature range. The kit contains a M-MLV reverse transcriptase with extended thermostability and half-life and contains proprietary mutations for reduced RNase H activity.

Kit Components:
  • Enzyme Mix: 100 µl
  • 2X Reaction Mix: 500 µl
  • Oligo dT20 Primer (50 µM): 50 µl
  • Random Hexamer Primer (50 ng/µl): 50 µl
  • Nuclease-Free Water: 1 ml

Materials Required But Not Provided:
  • Vortex Mixer
  • Microcentrifuge
  • Pipettes and Pipette Tips
  • PCR Tubes
  • Ice Water Bath
  • Temperature-Controlled Water Bath (or equivalent)


Target cDNA Synthesis Kit
Tested Applications PCR
Storage Store at -20 °C for up to 18 months. Avoid repeated freeze/thaw cycles.
Buffer Enzyme: 20 mM Tris-HCl, pH 7.4, 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, 0.01% NP-40 and 50% glycerol.
Concentration 200 U/µl
Availability Shipped within 10-15 working days.
Note This product is for research use only.
Directions for use Assay Procedure:

  1. Mix and heat the RNA Template, 2X Reaction Mix, and Primers to 65 °C for 5 minutes. Allow the RNA Template to stand in an ice bath for at least 1 minute.
  2. Add the following components to a PCR tube on ice:

    Component Volume
    RNA Template Variable
    10 pg - 2 µg total RNA or 10 pg - 500 ng mRNA)
    2X Reaction Mix10 µl
    Primers1 µl
    Enzyme Mix2 µl (200 U)
    RNase Inhibitor1 µl (20-40 U)
    Water Variable, up to 20 µl
    Total Volume 20 µl

  3. Mix by gently pipetting up and down.
  4. Close the lid on the tube and incubate in a temperature-controlled water bath at 55 °C for 50 minutes for the extension step.
    Note: the optimal temperature for extension is likely between 42-60 °C.
  5. Incubate the tube at 70 °C for 15 minutes to inactivate the Reverse Transcriptase before amplification.

Notes:
  • Avoid cross-contaminating the RNA template (total RNA, synthetic RNA transcript, poly(A) + mRNA) with DNA.
  • Recommended primers (per 20 µl reaction):
    • 2.5 µM of oligo(dT) - anneal to 3'-poly(A) + mRNA
    • 2.5 ng/µl of random primers - anneal at non-specific sites of RNA templates
    • 2 µM of gene-specific primers
Research Articles on cDNA Synthesis Kit


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