NGS RNA Second Strand Synthesis Module

The NGS Second Strand Synthesis Module contains the enzymes and buffers required to convert first strand cDNA into double stranded cDNA. The fast, user-friendly workflow has minimal hands-on time and is compatible with upstream first strand cDNA synthesis.

Applications

• RNA-Seq Library Construction.

Protocol 

1.     In a nuclease-free 0.2 mL PCR tube, mix the following components on ice:

         First Strand Synthesis Reaction Buffer            4 μL
         First Strand Synthesis Enzyme Mix                 1 μL
         Nuclease-free Water                                        4 μL
        Total volumes                                                   20 μL

2.     Mix by pipetting gently up and down.

3.     Incubate the samples in a preheated thermal cycler (with the heated lid set to 100°C):

       10 minutes at 25°C
       30 minutes at 68°C
       Hold at 4°C

4.     Place the tube on ice.

5.     Proceed directly to second strand synthesis using NGS RNA Second Strand Synthesis Module (Cat no. E118).