FastDigest BamHI

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	For each of the three gRNA expression cassettes, set up a restriction endonuclease digestion reaction as described below. Digest separately the gRNA expression segment gRNA1 with EcoRI and BamHI, gRNA2 with BamHI and NcoI, and gRNA3 with NcoI and NotI. Incubate the digestion mixture at 37 °C for 30 min. Purify the DNA from the digestion mixture with the Promega gel and PCR clean-up system. We show digestion of gRNA1 expression cassette from Step 12 as an example. Digest gRNA2 and gRNA3 with the corresponding enzymes in the same manner.

Protocol tips

For each of the three gRNA expression cassettes, set up a restriction endonuclease digestion reaction as described below. Digest separately the gRNA expression segment gRNA1 with EcoRI and BamHI, gRNA2 with BamHI and NcoI, and gRNA3 with NcoI and NotI. Incubate the digestion mixture at 37 °C for 30 min. Purify the DNA from the digestion mixture with the Promega gel and PCR clean-up system. We show digestion of gRNA1 expression cassette from Step 12 as an example. Digest gRNA2 and gRNA3 with the corresponding enzymes in the same manner.

Publication protocol

For each of the three gRNA expression cassettes, set up a restriction endonuclease digestion reaction as described below. Digest separately the gRNA expression segment gRNA1 with EcoRI and BamHI, gRNA2 with BamHI and NcoI, and gRNA3 with NcoI and NotI. Incubate the digestion mixture at 37 °C for 30 min. Purify the DNA from the digestion mixture with the Promega gel and PCR clean-up system. We show digestion of gRNA1 expression cassette from Step 12 as an example. Digest gRNA2 and gRNA3 with the corresponding enzymes in the same manner.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for FastDigest BamHI below.

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