Notebook

Experiment Overview

Here we will include a day by day entry for our project. We need to make sure we atleast include the days we did which experiments but more information would be good too. Note who participated in what should be included.

June 5

LB and LB Kan Plates were poured.

June 6

Re-streaked Ecoli BL21, DH5α, and DH5α/ pET-28a from the stocks we were provided.

June 7

Liquid cultures of DH5α, DH5α/ pET-28a and BL21 were inoculated and grown for 48 hours

June 9

Glycerol stocks were prepared from the liquid cultures

June 14

Ordered frc and oxc Full length Sequences

June 19

Ordered gBlock fragments of frc and oxc for use in a gibson assembly

June 20

Ordered original PCR primers to add PstI to pET-28a

June 21

A half sleeve of each chloramphenicol and Kanamycin plates were poured, DH5α/pET-28a was re-streaked, and a DH5α liquid overnight culture was prepared.

June 22

DH5α competent cells were made. Experiment failed at the final step due to forgetting the glycerol before putting cells in liquid nitrogen.

June 25

Ordered primers to add frc and oxc ends to pET-28a(PstI).

July 5

Miniprep DH5α/pET-28a, and PCR to add PstI to pET-28a.

July 6

Gibson Assembly transformation.

July 10

Made LB broth and an agarose gel.

July 12

Re Streaked BL21 (onto LB) and DH5α/pET-28a (onto LB Kan).

July 13

Overnight liquid cultures were prepared of BL21 and DH5α / pET-28a in 4 mL LB broth in plastic culture tubes. Kanamycin was forgotten in pET-28a culture. Ordered Primers for frc and oxc genes without RE end sites.

July 14

Prepared BL21 Competent Cells, Miniprep of DH5α / pET-28a.

Insert Day here