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Revision as of 22:58, 26 September 2013
crRNA-Design
This tool helps you to design a crRNA-insert for pX334a. The oligos contain overhangs which fit to the BbsI-overhangs by pX334a.
For repression of gene transcription by targeting the coding sequence
it´s crucial to target the non template (= coding) DNA strand.
Therefore the oligos must be designed as
follows:
- Search at your desired target sequence for a CCN (reverse complement of the PAM sequence) at the coding strand.
- Extract the following (3') 30 nucleotides.
- Extract the reverse complement.
- Add AAAC at the 5' end and GT at the 3' end. This will be your fist oligo.
- Take the sequence from step 2 and add TAAAAC at the 5' end. This will be your second oligo.
