....The actual oligo you should order is:5'-CACCGN...N-3'3'-CN...NCAAA-5'You need to add the CACC and AAAC...Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A...."CRISPR", short-hand for Clustered Regularly Interspaced Short Palindromic Repeats, in Pubmed returned...want to use double nickase, you could express two spacers and use PX335 to express the Cas9n (nickase).
The...PAM sequence to the oligo when cloning my target spacers into the PX330 vector backbone?
A2: There is no...usually adds it when they order the oligo. If your spacer sequence starts with a 'G', you naturally have ... compared to where the target site sequence (protospacer+PAM) is located? If there are other PAMs in the...