New England Biolabs
HiScribe T7 High Yield RNA Synthesis Kit
E2040S
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The WNT pathway is essential for embryonic development and adult tissue homeostasis. Deregulation of this pathway results in cancers of the stomach, intestine, breast, etc. I am interested in studying the molecular mechanism of oncogenic WNT signal transduction to identify potential targets for therapeutic intervention of colon cancer. The goal of this experiment is compare the editing efficiency of sgRNA purchased from a commercial vendor to that of a homemade version using the NEB HiScribe T7 High Yield RNA Synthesis Kit in CRISPR/Cas9 RNP format.
RNA synthesis for CRISPR/Cas9 application
dsDNA obtained by PCR amplification (template)
Follow manufacturer's instructions.
I routinely test several CRISPR guide RNA sequences to pick guides with the best INDEL efficiency. This can get expensive if the guide has to be obtained from a commercial vendor. So, I had tested this RNA synthesis kit from NEB for my CRISPR/Cas9 RNP experiment. The editing efficiency using this homemade sgRNA was similar to the one from a commercial vendor.
N/A
16 h incubation yields 70-100 µg of high quality RNA.
None.
Cost-effective option for CRISPR sgRNA synthesis.