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The crRNA and tracrRNA were received as pellets, which were individually resuspended in Duplex buffer (IDT) to form 200 μM stocks. Stocks were stored at −80°C before use.
The crRNA was annealed with the tracrRNA to form the gRNA by mixing each crRNA of the set separately with an equal molar amount of tracrRNA and diluting to 57 μM in Duplex buffer. This was usually: 1 μL crRNA 200 μM; 1 μL tracrRNA 200 μM; 1.51 μL Duplex buffer, heated to 95°C for 5 min, then cooled on ice.
Copyright and License information: ©2021, Kroll et al
This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
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