Team:Caltech/Week 2

From 2011.igem.org


Caltech iGEM 2011



Home

Project

Data

Parts

Team

Notebook

Biosafety

Human Impact

References

Support

June 19

Collection of soil and water samples from LA River

June 20

Lab walkthrough
Setting up lab, stock supplies, preparation of LB-amp plates
Test extraction of DNA from BioBrick wells into cloning plates (K123000, K123001, K123002, K123003)
Initial enrichment cultures of collected samples on minimal media with BPA or 17a-ethynylestradiol

June 21

MoBio Powermax Soil kit extraction of DNA from collected samples
Initial enrichment cultures of samples in minimal media with nonylphenol or DDT
Geneious and primer design tutorial

Results

MoBio Extraction

Sample Concentration (ng/µl)
1
4.5
2
4.1
4
0.0
5
0.7
6
13.7
7
5.8
9
58.7
10
11.4

Locations 3 and 8 were omitted from the MoBio Powermax Soil Extraction procedure because they were liquid samples.

June 22

Miniprep and sequencing of selected BioBricks (BBa_K123000, BBa_K123001, BBa_K123002, BBa_K123003)
Preparation of competent cells

Results of OD measurements:

Part Concentration (ng/µl)
K123000
326.6
K123001
293.8
K123002
152.3
K123003
402.6

June 23

Transformation of T7 polymerase (K145001), mCherry (J06702]), Lac Promoter (R0010]), and double terminator (B0014 and B0015) biobricks for test sequences for BisA and BisB degradation genes (K123000 and K123001)
Retested competent cells made June 22nd with K123001 and pUC
Transfered 0.5 mL aliquots of BPA and 5mL 17a-ethynylestradiol cultures from June 20 to new tubes of minimal media
Designed primers for test sequence of BisdA (pNT001)
Designed primers to continue sequencing of human ER (K123003)

We determined that sequences for K123000 and K123001 have correct protein coding but different codons than in the sequences shown online

June 24

Transformation of Tet repressible promoter R0040
Preparation of LB, LB (no antibiotic) plates, and SOC media
Transfer of 5 mL DDT and nonylphenol cultures from June 21 to new tubes of minimal media
Design of reverse primer for continued sequencing of human ER (K123003)
Update Wiki with protocols we have been using

We determined that our competent cells are functional have a cfu/ug DNA of 6.6 x 10^6.

June 25

Take out plates from June 24


Retrieved from "http://2011.igem.org/Team:Caltech/Week_2"