check by cut NdeI+EcoRI or NdeI+HindIII

after ligation and transformation we get screened colonies

and we should check the plasmid we picked by miniprep and digestion to tell if it succeeds.

 

 

 

 

2. EcoRI + NdeI

 

 

 

 

 

 

 

 

 

 

 

 

 

 

3. EcoRI + NdeI

4. EcoRI + NdeI

 

1.to watch their band shifting and to digest it by EcoRI + NdeI

 

 

 

 

5. EcoRI + NdeI ----dM9 3.4 are correct ! (by another PCR check not listed)

 

6. dRG check by EcoRI + NdeI

 

 

 

 

PCR check

 

 

 

 

 

 

 

 

 

 

6.repeat EcoRI + NdeI

 

 

 

 

 

 

 

 

 

 

6. cut by NdeI + HindIII

PCR check & HindIII + NdeI (check direction) dRG 1& 8 are correct!