Team:Groningen/Notebook/3 August 2009
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+ | 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). | ||
{{Team:Groningen/Notebook/Day/Footer}} | {{Team:Groningen/Notebook/Day/Footer}} |
Revision as of 14:49, 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).
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