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transformation of MCS-pcc009and pcc010

Smear the transformation onto Cm+LB plate

plx007,008,009 positive transformation

Pick colonies

colony PCR of RD2-pGEM-T

using Primer M13F/R

T1TE-pcc010 miniprep

double digestion using KpnI/XhoI and M buffer pcc010 as the negative control

Miniprep of plx007/008/009

using XhoI/SalI to double digest in H buffer

digestion of vector 7-6, 8-6, 9-5

plasmid 2uL

Buffer H 10uL

XhoI/SalI 2uL each

H2O 84uL

total 100uL

fragment lacZa-XS 16uL

buffer H 2uL

SalI 2uL

H2O 70uL

total 100uL