SHEATECH, INC
Gene Expression and Proteomics

Other Projects

DNA Assembly Cloning

 

T5 exo nuclease removed 5’end of double strand DNA

Single stand DNA annealed

Pfu DNA polymerase filled the gaps

Tag DNA ligase sealed the nicks at 50oC

Direct transform to DH5a and/or transfect to sf9 insect cells


 


 

 

 

One step isothermal assembly reaction of linearized transfer vector and PCR fragments. From lane2 to lane5, each sample has one linearized transfer vector and one PCR product (PCR 1). From lane6 to lane9, each sample has one linearized transfer vector and two PCR products (PCR 1 & PCR 2). Both lane3 and lane7 are negative control samples. Lane5 sample is the negative control of lane9 sample. The DH5a transformed results were expected and colonies were confirmed by mini preparation and restriction enzyme analysis.