Team:Munich/cfp gRNA.html

Generation of gRNA (the DNA for the transcription)

2018/05/25

Participants:	Keno Eilers
Protocol:	gRNA production
Notes:	Work at RNAse free bench.
Results:	Coding and non-coding part of digestion site 1, 2 and 3 are annealed.

Generating MS2 RNA and gRNA

2018/05/28

Participants:	Keno Eilers
Protocol:	gRNA production, Phenol Chloroform precipitation
Notes:	No notes.
Results:	DNA has been used for MS2production and gRNA for cpf digest.

Cpf1 digest -gRNA production

2018/05/29

Participants:	Keno Eilers
Protocol:	Cpf1 digest, Urea gel
Notes:	No notes.
Results:	No gRNA visible.

Cpf1 digest

2018/06/06

Participants:	Keno Eilers
Protocol:	Cpf1 digest, Agarose gel
Notes:	No notes.
Results:	No bands visible.

Generating new gRNA

2018/06/14

Participants:	Keno Eilers
Protocol:	gRNA production, Phenol Chloroform precipitation
Notes:	Used DNAse I
Results:	DNA has been used for Urea Page gel

gRNA generation and Urea Page Gel

2018/06/18

Participants:	Keno Eilers
Protocol:	gRNA production, Urea gel
Notes:	No notes.
Results:	gRNA1 620 ng/µl gRNA2 591 ng/µl

Cpf1 digest with gRNA

2018/06/26

Participants:	Keno Eilers
Protocol:	Cpf1 digest, Agarose gel
Notes:	No notes.
Results:	No bands, T7 DNA not clean enough.