![]() The 1-base overhangs produced by BmrI may be hard to ligate.The plasmid vector platform is the most commonly used vector for the expression of the versatile CRISPR-Cas technique and the promoter is a crucial element for the expression vector, thus profiling the impact of the promoters on CRISPR editors provides the basic information for the gene-editing toolkits and can be a guideline for its design. Sticky ends from different BglI sites may not be compatible.Ī C T G G G ( N ) 4 N T G A C C C ( N ) 4 G C C N N N N N G G C C G G N N N N N C C G Sticky ends from different BstAPI sites may not be compatible.īsaHI is typically used at 37☌, but is even more active at 60☌.Įfficient cleavage requires at least two copies of the NarI recognition sequence.Įfficient cleavage requires at least two copies of the PluTI recognition sequence. G C A N N N N N T G C C G T N N N N N A C G Prolonged incubation with NdeI may lead to removal of additional nucleotides. Sticky ends from different PfoI sites may not be compatible.
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