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: 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. 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 Mix 10 µl Primers 1 µl Enzyme Mix 2 µl (200 U) RNase Inhibitor 1 µl (20-40 U) Water Variable, up to 20 µl Total Volume 20 µl Mix by gently pipetting up and down. 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. 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

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