"
Team:Groningen/Notebook/3 August 2009
From 2009.igem.org
(Difference between revisions)
(→Dry) |
(→Dry) |
||
| Line 49: | Line 49: | ||
==Dry== | ==Dry== | ||
We (Jasper and Klaas) worked on deriving a method for estimating K and V<sub>max</sub> values based on combined import/export measurements. So far the application of this method on figure 3A of [[Team:Groningen/Literature#Kostal2004|Kostal2004]] has only yielded nonsensical results (K and V both negative and K (close to) zero if they are constrained to be positive). | We (Jasper and Klaas) worked on deriving a method for estimating K and V<sub>max</sub> values based on combined import/export measurements. So far the application of this method on figure 3A of [[Team:Groningen/Literature#Kostal2004|Kostal2004]] has only yielded nonsensical results (K and V both negative and K (close to) zero if they are constrained to be positive). | ||
| + | |||
| + | we used the data of the plasmid, in the e. coli we used the chromosal ArsR, to see the difrence between the genes I have the data alignment with clustalW | ||
| + | |||
| + | |||
{{Team:Groningen/Notebook/Day/Footer}} | {{Team:Groningen/Notebook/Day/Footer}} | ||
Revision as of 15:01, 3 August 2009
 |
|---|
Wet
GVP Cluster
Over Night Cultures
Over night cultures in 4mL LB-amp100 or LB-kan50 medium were prepared from the following glycerol stocks:
- → E.coli TOP10 pSB3K3-high const. promoter (kan.)
- → E.coli TOP10 pSB3K3-med. const. promoter (kan.)
- → E.coli TOP10 pSB3K3-low const. promoter (kan.)
- → E.coli TOP10 pSB1AC3-high const. promoter (amp.)
- → E.coli TOP10 pSB1AC3-med. const. promoter (amp.)
- → E.coli TOP10 pSB1AC3-low const. promoter (amp.)
and put in the 37°C waterbath at 200 rpm.
Transporters
DONE GlpF → pick colonies and use for overnight culture. (2 colonies in 6 mL LB-amp medium)
Metal Accumulation
TODO MBP
- Plasmid isolation (cells can be found in the freezer Jolanda)
- PCR of MBP
- PCR Mix
| 12,5 uL | Phusion 2x MM |
| 1 uL | MBP Fw |
| 1 uL | MBP rev |
| 0.5uL | MBP plasmid |
Vectors
Dry
We (Jasper and Klaas) worked on deriving a method for estimating K and Vmax values based on combined import/export measurements. So far the application of this method on figure 3A of Kostal2004 has only yielded nonsensical results (K and V both negative and K (close to) zero if they are constrained to be positive).
we used the data of the plasmid, in the e. coli we used the chromosal ArsR, to see the difrence between the genes I have the data alignment with clustalW
|
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
