IGEM:Peking/2007/Switch-Notebook/2007-8-12
rescue by precipitation of fragment after digestion
electrophoresis result to show that SD2, SD3 and Prec successfully ligated.
Colony PCR of T1T2-pcc010
easy Taq 0.1uL
Pkan-F 0.5uL
pcc-test-R 0.5uL
Buffer 2uL
dNTP 2uL
ddH2O 15uL
We did choose some colonies that seems to be right according to the PCR band after electrophoresis.
Miniprep and double digestion test
T1T2-Pcc010
plasmid 3uL
SacII 0.5uL
T buffer 2uL
BSA 2uL
ddH2O 12.5uL
T1TE-T1T2-pcc010
plasmid 3uL
SalI 0.5uL
XhoI 0.5uL
H buffer 2uL
H2O 14uL
SD2 and SS2
plasmid 3uL
SalI 0.5uL
XhoI 0.5uL
H buffer 2uL
H2O 14uL
total 20uL system
re-PCR
lac, rec, sulA, SS1-3,SD1-3
Some of the fragment did appear this time after we change some data in PCR processing.
test of enzymatic sites on GFP-plx007
plasmid 10uL
XhoI/SalI 0.5uL each
buffer H 2uL
H2O 7uL
or
Plasmid 4uL
XbaI 0.5uL
buffer M 2uL
H2O 13.5uL
total 20uL system
ligation of SD2, SD3 and Prec with SDY-EGFP-plx007
ligation of SS1, SS2 and SS3 with SDY-EGFP-plx007
praparation of LB medium with Cm
miniprep of T1T2-pcc010
single digestion to test if the plasmid we isolated it right or not
digestion test of T1TE-T1T2-pcc010
plasmid 2uL
H buffer 10uL
Trition X-100 10uL
BSA 10uL
NotI/XhoI 2uL each
add ddH2O to a total of 100uL system
