07/23/2014

Today we have performed a Plasmidial DNA extraction of JM110 and E0840. We have also checked the electrophoretic profile of all our samples, to keep it for when it’s needed.

Eletrophoretical profile display:

1: E0840 extracted with DH5α

2: K346004extracted with DH5α

3: K346004 extra with Kit of JM110

4: E0840 extracted of JM110 electrocompetent (Colônia 1)

5. E0840 extracted JM110 eletrocompetent (Colony 2)

6. E0840 extracted of JM110 electrocompetent (Colony 13)

7. E0840 extracted with Kit of JM110 eletrocompetent (Colony 13)

8. E0840 extracted with Kit of JM110 chemical competent

Today we have also digested all the samples above with XbaI and PstI, to analyse the cut and the size of the cut fragment. In theory, Xbal is not supposed to cut its restriction sites from the samples extracted of DH5α, taking into account that it’s not dam negative. We will see if we can remove the dam methylation from our DNAs so that we could work in our constructions!!!!

Digestion system (XbaI and PstI) 1 hour at 37°C

- 18ul of Water

- 1ul of DNA

- 1ul of Endonucleases

- 5ul of NEB 3 buffer

Eletrophoretical display of the digested samples: