Editing 3131 pCEL0-HA-VL-mIgKappa

Subclone of 3131:

Restriction enzymnes:BssHII+XhoI

Time:30min

Buffer:Fastdigest

result:good, combine and do ligate

[[File:Subclone of new 3131.JPG|RTENOTITLE]]

'''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

[[File:Miniprepof 3131 3167 3161.JPG|RTENOTITLE]]

'''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

[[File:Redo miniprep of 3131 3161 3167.JPG|RTENOTITLE]]

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'''

[[File:Miniprepdigstion3131.JPG|RTENOTITLE]]

'''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'''

[[File:IMG 3621.JPG|RTENOTITLE]]

Date:3/19/2018

'''Redo Subclone'''

'''3131''':BssHII+XhoI

Buffer:Cutsmart

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

[[File:IMG 3626.JPG|RTENOTITLE]]

'''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.'''



[[File:IMG 3637.JPG]]
[[Category:Cloning]]