3131 pCEL0-HA-VL-mIgKappa

Subclone of 3131:

Restriction enzymnes:BssHII+XhoI

Time:30min

Buffer:Fastdigest

result:good, combine and do ligate

Miniprep digestion of 3131_3161_3167

Name length Enzymen Buffer

3131:1-4 342 BssHII+XhoI Cutsmart

3161:5-8 225 NdeI+XhoI Cutsmart

3167:9-12 397 BsrGI+NheI NEB 2

Time:40min

Redo miniprep of 3131_3161_3167

3131:1-4 BssHII+XhoI 342 Cutsmart

3161:5-8 NdeI+XhoI 225 Cutsmart

3167:9-12 BsrGI+NheI 397 NEB 2

Time:40min

Date:3/15/2018

MiniPrep digestion of 3131

Enzymes:BssHII+XhoI

Buffer:Cutsmart

Time:40min

Result:2,3,4,seems good, do sequencing,select #3

Miniprep of 947_947pMiniT2_3131_3152

947:1-4 XbaI+NcoI-HF 1394 Cutsmart

947pMiniT2:5-8 ECoRI Cutsmart

3131:9-12 BssHII+XhoI 342 Cutsmart

3152:13-16 BsrGI+KpnI 371 NEB 2

Result: sent lane 2, lane 16 to do sequence

Date:3/19/2018

Redo Subclone

3131:BssHII+XhoI

Buffer:Cutsmart

Results:cut out the middle band of insert, and cut out the vector.

Date:3/21/2018

Miniprep of 3165pMiniT2_3165_3131

3165pMiniT2:1-5 EcoRI

3165:6-10 BsrGI+NdeI

3131:11-15 BssHII+XhoI

Time:30min

Result:3165pMiniT2(1-5) have bands,3131(12-15)have bands.redo 3165 digestion.