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G3337 SweScript All-in-One First-Strand cDNA Synthesis SuperMix for qPCR (One-Step gDNA Remover)

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G3337 SweScript All-in-One First-Strand cDNA Synthesis SuperMix for qPCR (One-Step gDNA Remover)

This product contains 5×Ready-to-use master mix (5×SweScript All-in-One SuperMix for qPCR) and gDNA remover reagents, can remove genomic remnants of the template while the reverse transcription reaction is performed.The kit contained 5×SweScript All-in-One No-RT Control SuperMix as a reverse-transcriptase-free control to determine whether the qPCR template was derived from cDNA.The master mix contains all reagents required for the reverse transcription reaction(SweScript RT, RNase Inhibitor, Random Primer, Oligo(dT)18 Primer, dNTPs and the optimized buffer system). SweScript RT is a mutant reverse transcriptase obtained by in vitro evolutionary screening and is characterized by high thermal stability and processivity. gDNA Remover contains a thermosensitive DNase that is 30 times more active than DNase I and acts only on dsDNA and is susceptible to inactivation under high temperature conditions.

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Product name Product code No. of reactions
SweScript All-in-One First-Strand cDNA Synthesis SuperMix for qPCR (One-Step gDNA Remover) G3337-50 50
G3337-100 100

 

Technical Specification

Storage and Handling Conditions

Wet ice bag transportation; Stored at -20ºC temperature, valid for 12 months.
 

Assay Protocol / Procedures

1.Synthesis of the first strand cDNA and Genome removal

(1) Preparation reaction system (recommended 20ul reaction system) :

Component Volume

5×SweScript All-in-One SuperMix for qPCR

4 μL

gDNA Remover

1 μL

Total RNA/mRNA

0.1 ng-5ug / 10 pg-0.5ug

Nuclease-Free Water

Add to 20 μL

(2) Gently mix and centrifuge;

(3) Reverse transcription program setting:

Temperature Time

25ºC

5 min

42ºC

15-30 min

85ºC

5 s

 

Note:

  1. Please wear disposable gloves during operation to avoid RNase contamination.
  2. Be sure to place 5×SweScript All-in-One SuperMix, 5×SweScript All-in-One No-RT Control SuperMix, gDNA Remover on ice.
  3. 5×SweScript SuperMix, 5×SweScript All-in-One No-RT Control SuperMix, gDNA Remover contains high concentrations of glycerol and should be briefly centrifuged before use.
  4. For high GC or complex templates, the reverse transcription temperature can be raised to 65ºC.
  5. If a negative control is required, just replace the equal volume of G3337-1 5×SweScript All-in-One SuperMix for qPCR in the reaction system with G3337-4 5×SweScript All-in-One No-RT Control SuperMix for qPCR.
  6. The reverse transcription products can be stored at -20ºC for a short time. If long-term storage is needed, it is recommended to store them at -80ºC after packing to avoid repeated freeze-thaw cycles.
  7. If the subsequent qPCR primers are designed across exons, the genome removal step can be omitted.
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