We have compared the performance of the Template Switching RT Enzyme Mix and Maxima H Minus Reverse Transcriptase (Thermo Fisher Scientific). The cDNA libraries made with the NEB Template Switching RT Enzyme Mix contain at least 10% more transcripts than those made with Maxima H Minus supplemented with 7.5% PEG as described in the mcSCRB method. In addition, the NEB libraries exhibited improved RNA base distribution metrics as listed below:
RT Options |
% Exon |
% rRNA |
% Intron |
% Intergenic |
---|---|---|---|---|
NEB |
77.8±0.3 |
2.1±0.3 |
10.3±0.0 |
9.7±0.0 |
mcSCRB |
50.0±1.8 |
24.8±1.8 |
9.8±0.2 |
15.4±0.2 |
1 Bagnoli, JW. et al. (2018) Nat Commun. 2937-2944 .