Type or paste the protospacer, 20 nt, 5′ to 3′, without the NGG PAM. Everything else follows as you type, and the sheet stays in this browser for next time.
| Target gene | # | Protospacer | Cloning oligos | Status |
|---|
One guide per line. Separate fields with a tab, comma or semicolon: the longest field that reads as DNA is taken as the sequence, the first other field as the gene. A line with no sequence in it is reported, not guessed at. Rows already here are kept.
Name and sequence, tab separated, in the order shown above.
Forward oligo = caccg + protospacer. Reverse oligo = aaac +
reverse complement of the protospacer + c.
BsmBI (Esp3I) leaves the same overhangs on lentiCRISPRv2, lentiGuide-Puro and lentiCRISPRv1, so one oligo pair serves all three. Zhang lab protocol rev20140722. pX330 takes the same oligo pair but is cut with BbsI (BpiI), not BsmBI — check which enzyme your backbone needs before digesting.
The NGG PAM is never part of the oligos. Standard de-salted synthesis is enough; resuspend to 100 µM in sterile water or TE.
This reproduces the lab spreadsheet exactly, including the caccg prefix on
guides that already begin with G. If your vector convention drops that extra g, edit the
sequence by hand before ordering.
Nothing here leaves your computer. The sheet is stored in this browser only.