Team:Bologna/Lab-Notebook

From 2009.igem.org

Revision as of 08:41, 21 August 2009 by Marco.cavina (Talk | contribs)

DnaMenu.png
HOME PROJECT TEAM SOFTWARE MODELING LAB-NOTEBOOK SUBMITTED PARTS HUMAN PRACTICE


Contents

Week 1: from 07/20/09 to 07/24/09

Trasfomation of PSB1K3, PSB3K3, PSB1A2, PSB4A3(2007). Only the last one seems to work. We also use PSB1A2.

Trasformation and Amplification of A, B, C:
A) PSB3K3(LMC) + 1429 + RBS + LACI
B) PSB1A2(HC) + 2547 + Ox + RBS + GFP + T
C) PSB3K3(LMC) + 1429 + RBS + GFP + T

Working (using DH5alfa) on:
A + B ---> Evaluation GFP fluorescence imaging comparing with the rRBS one
B ---> Evaluation GFP fluorescence imaging
C ---> Evaluation GFP fluorescence imaging
B+C ---> Evaluation ΣGFP fluorescence imaging

Up

Week 2: from 07/27/09 to 07/31/09

Sequences suspension to have 100 μM conc. (EP digestion and trasformation in a HCN plasmid with AMP)

DNA sequences Plasmid
H2O mQ == 4.5 μM H2O mQ == 19.5 μM
Sequence == 20 μM DNA (miniprep) == 5 μM
Buffer == 3 μM Buffer == 3 μM
BSA == 0.5 μM BSA == 0.5 μM
EcoRI enzyme == 1 μM EcoRI enzyme == 1 μM
Pst1 enzyme == 1 μM Pst1 enzyme == 1 μM
TOT: 30 μM TOT: 30 μM


Gel Run:

1.CIS
2.TRANS 4
3.TRANS 7
4.PSB1A2

(Figure)


Up

Week 3: from 08/3/09 to 08/7/09

Amplification of CIS, TRANS 4 and TRANS 7 using PSB1A2. Injection. Unexpeted results from the gel run (3000, 2200, 100 bp instead of 2100, 2000, 100 bp). Done the ligation again whit the some bad results. Done some test fluorescence measurements using GFP in plasmids with different copy number.


Up

Week 4: from 08/10/09 to 08/14/09

HOLIDAY!!


Up

Week 5: from 08/17/09 to 08/21/09