"
Team:ESBS-Strasbourg/17 July 2008
From 2008.igem.org
(Difference between revisions)
(→General) |
(→DryLab) |
||
| (4 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
[[Team:ESBS-Strasbourg/Notebook | < back]] | [[Team:ESBS-Strasbourg/Notebook | < back]] | ||
== DryLab == | == DryLab == | ||
| + | |||
| + | michi: | ||
| + | *found paper PMID 18385157 which mutates TATA box and quantifies transcription :-) (King of the day) | ||
=== Modeling === | === Modeling === | ||
| Line 17: | Line 20: | ||
</pre> | </pre> | ||
| - | *The new BioBricks arrived | + | *The new BioBricks arrived so we made an overnight culture of, in LB(Amp): |
| + | **p1010 in pSB1A3 | ||
| + | **BBa_c0012 (lacI) | ||
| + | **BBa_c0051 (lamda cI) | ||
| + | **BBa_j63001 (EYFP) | ||
| + | *We plated p1010 in pSB3K3 on LB Kan | ||
| + | |||
*Digestion of the plasmid of the BBa_J63002(?) clone with XbaI and SpeI | *Digestion of the plasmid of the BBa_J63002(?) clone with XbaI and SpeI | ||
**1x double digestion | **1x double digestion | ||
**1x step by step digestion | **1x step by step digestion | ||
| + | **result: | ||
| + | ***digestion seem to have work out in both cases | ||
| + | ***but the gel shows a high contamination with low molecular RNA, so we know now that RNase is crutial in the eluation buffer | ||
| + | *** the identity of the clone is still not verifified, because the size small fragement could be determinated | ||
| + | *** but as the new BioBricks arrived today, there is no longer an interest to go on working on this clone | ||
== General == | == General == | ||
Latest revision as of 09:25, 20 July 2008
Contents |
DryLab
michi:
- found paper PMID 18385157 which mutates TATA box and quantifies transcription :-) (King of the day)
Modeling
WetLab
- Results Transformation
- no clones for any of the three BioBricks
- clones for the LexA-BD plasmid --> useable for Miniprep
- counting:
- dh5a (Mariel) + pxj - KCM : 10^5 cfu/µg - dh5a (Mariel) + pxj - HS : 4*10^4 cfu/µg - dh5a + pxj - HS : 1.5*10^5 cfu/µg - Top10 + pxj - HS : 3.2*10^5 cfu/µg - db3.1 + pxj - HS : 3*10^4 cfu/µg
- The new BioBricks arrived so we made an overnight culture of, in LB(Amp):
- p1010 in pSB1A3
- BBa_c0012 (lacI)
- BBa_c0051 (lamda cI)
- BBa_j63001 (EYFP)
- We plated p1010 in pSB3K3 on LB Kan
- Digestion of the plasmid of the BBa_J63002(?) clone with XbaI and SpeI
- 1x double digestion
- 1x step by step digestion
- result:
- digestion seem to have work out in both cases
- but the gel shows a high contamination with low molecular RNA, so we know now that RNase is crutial in the eluation buffer
- the identity of the clone is still not verifified, because the size small fragement could be determinated
- but as the new BioBricks arrived today, there is no longer an interest to go on working on this clone
General
- Modeling meeting with Mr. Haich
Quote of the day
Marius loves Michis Kidneys!
