|For goldengate reaction, there is a low and a high concentration mixture available for T4 ligase. For single inserts, the low concentration is just fine, but the follow-up article on golden gate cloning (PMID: 19436741) found that the high-concentration T4 was better for multiple insert cloning.|
pKLV2-U6gRNA5(BbsI)-PGKpuro2ABFP-W from Addgene has not yet been reviewed for this experiment
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3 years ago
Author: Milena Alexeyeva
I would like to excise a large strand of DNA and insert a new one using CRISPR. My problem is that my strand will be a little over 1kb and I am not sure if this is going to be a limiting factor. Also, how long should the homology arms be for a region of this size?
Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Human - Deletion KAT2A using pKLV2-U6gRNA5(BbsI)-PGKpuro2ABFP-W from Addgene.
Download the product protocol from Addgene for pKLV2-U6gRNA5(BbsI)-PGKpuro2ABFP-W below.Download manufacturer protocol
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