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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


plasmid 3uL

SacII 0.5uL

T buffer 2uL


ddH2O 12.5uL


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


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


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