Matt Gethers/CRI, Thailand/Labwork/Generating the HmgR Mutant/Week of 7.13.08

7.14.08

To Do:

• Ligate the HmgR downstream fragment (p'So #4) into pKn002 and HmgA upstream fragment (p'So #2) into pKn001.

Summary:

Performed the ligations today and placed the products in the -20 for the day. I did a transformation in the evening.

7.15.08

To Do:

• Make colony suspension of transformants, start 5 ml cultures, do PCR.

Summary:

One colony from the pKn003.L2 transformation looks promising (See results here). I took the culture I started this morning from colony 5, made a glycerol and miniprepped. I sent the prep for sequencing. I should probably sequence another round of colonies from pKn003.L2 so I have more than one candidate to work with. I need to check the efficiency of my digest with HindIII/SphI and try the ligation and transformation again.