Team:Munich/cas13a.html

Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA

2018/04/24 - 2018/04/26
Participants: Katja Neishsalo
Protocol: Restriction Digest, Gibson Ligation, Chemical Transformation
Notes: Digest of gRNA-EndoA gBlock with NheI-HF and consequent heat inactivation 80°C. Samples were gibson ligated into p2ct_his_MBP_LBU_c2c2 and E.Coli NEB Turbo was transformed. Sample DNA was preped and digested to verify the correctness.
Results: Restriction digest showed false results; Thomas (supervisor) and Katja decided to redo the experiment

Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA

2018/05/16
Participants: Katja Neishsalo
Protocol: Restriction Digest, Gibson Ligation, Miniprep, Chemical Transformation
Notes: 2x chemical transformation (turbo, dh5a) 1x electroporation (dh5a)
Results: Some colonies. Sample DNA was prepped and digested to verify the plasmid. Restriction digest gave no correct bands for the plasmid. We suspected that the Cas13a template is not good anymore.

Test digest of p2ct_his_MBP_LBU_c2c2_WT

2018/05/28
Participants: Katja Neishsalo
Protocol: Restriction Digest
Notes: SalI NheI;
Results: Restriction enzymes and template are OK (PIC)