Revision as of 12:22, 24 September 2012

Wageningen UR

week 10: 2 july - 8 july

TuYV

Monday:

A new transformation of last weeks ligation mixes into our electrocompetent 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 constrnct 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 constrnct 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 preperations

2nd – 5th July (Mark)

6th July

@@ Line 49: / Line 49: @@
 ----
+'''FPLC S400 preperations'''
+''2nd – 5th July'' (Mark)
+<ul>
+<li>Pouring and running of the S400 FPLC with the help of Serve Kengen.</li>
+</ul>
+''6th July''
+<ul>
+<li>FPLC with the S400 column</li>
+<li>1 run with 0.5 ml sample</li>
+<li>A very good separation with low concentration</li>
+</ul>
+----
 [[https://2012.igem.org/Team:Wageningen_UR/Journal/week9 previous week]]           [[https://2012.igem.org/Team:Wageningen_UR/Journal/week11 next week]]