Product Class: Kit

ProtoScript® II First Strand cDNA Synthesis Kit

Product Introduction

The ProtoScript® II First Strand cDNA Synthesis Kit is optimized mix for higher specificity and yield of cDNA.

  • Recombinant M-MuLV reverse transcriptase with reduced RNase H activity and increased thermostability (active up to 48°C)
  • Generates cDNA up to 10 kb
  • Supplied with Oligo-dT primers, Randomized Primer Mix, and nuclease-free water
Catalog # Size Concentration
E6560S 30.0 reactions
E6560L 150.0 reactions

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    Avoiding RNase Contamination

Protocols, Manuals & Usage

Protocols

  1. First Strand cDNA Synthesis Protocols (E6560)

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. How do I choose between LunaScript® RT SuperMix Kit and ProtoScript first strand cDNA synthesis kits?
  2. What is the difference between E6560 and E6300?
  3. What is the reaction temperature for E6560?
  4. Is RNaseH treatment required before PCR amplification?
  5. Can the cDNA products be used in real-time PCR analysis?
  6. What thermostable DNA polymerase can be used for PCR after cDNA synthesis?
  7. How do I choose between Induro Reverse Transcriptase and the different LunaScript and ProtoScript products?
  8. Why do I have low cDNA yields?
  9. How do I know whether my template RNA is of good quality?

Troubleshooting

Problem Suggestion
Low Yield of cDNA Check the integrity of the RNA by denaturing agarose gel electrophoresis (2). RNA should have a minimum A260/A280 ratio of 1.7 or higher. Ethanol precipitation followed by a 70% ethanol wash can remove most contaminants such as EDTA and guanidinium. Precipitation with lithium chloride can remove polysaccharides (2).
  Phenol/chloroform extraction and ethanol extraction can remove contaminant proteins such as proteases (2).
  Some target RNA may contain strong pauses for RT; Use random priming instead of d(T)23VN.
  Use sufficient amount of RNA.

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