week 10: 2 july - 8 july

Lab work

TuYV

Monday:

A new transformation of last week's ligation mixes into our electro-competent DH5 Alfa E. Coli was performed. The next day we checked the colony, there were many colonies on the positive control, however there were the same level of colonies on the sample and negative control. The transformation was contaminated.

Because there are iGEM illegal sites in our TuYV readthrough part, we designed a new construct named 4 and 4H, which were coat protein with part of the readthrough without illegal site and coat protein with part of the readthrough without illegal site with his-tag.

We tried to isolate and amplify 4 and 4H today, but we did not get any band on the agrose gel check

Tuesday:

Today we successfully PCR amplified the construct 4, which should be around 800bp.

Figure 1: Isolation of TuYV construct 4 from the TuYV genome

Wednesday:

Today we tried another time to PCR amplify the construct 4H and we got expected band on the gel check.

Figure 2: Isolation of TuYV construct 4H from the TuYV genome

DLS boundary experiment

6th July (Mark)

Growing CCMV for the temperature and pH stability experiment
Inoculated 200 mL of LB with BL21 that contains the IPTG inducible CCMV wild type monomer
Followed protocol of producing CCMV

7th – 10th July (Mark)

FPLC S400 preparations

2nd – 5th July (Mark)

6th July (Mark)