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'''Karla''' | |||
* Engineer other bacteria to photosynthesize (bacillus and e coli) | |||
* Find transformation system for Heliobacteria | * Find transformation system for Heliobacteria | ||
** Anerobic jar’s don’t work great, little room for plates | |||
*** Stack w/stick or paper towel roll to have more plates per jar | |||
** Heliobacter grow to slow/don’t grow, problems with temp | |||
*** Try top agar or pour agar | |||
'''Mike''' | |||
* Bacolovirus doesn’t want to express NELF, kills them? | |||
**Try using a signal peptide to target the proteins for secretion to get them away from the cells and hopefully decrease toxicity | |||
'''Cell-free expression''' | |||
* New cell-free expression system with dual rxn setup (separated by membrane). Synthesis rxn and feeder rxn for substrate to diffuse from and inhibitory products to fuse into. | |||
**Email Doug | |||
'''Wiki''' | |||
* Bede had previously had a good idea about collecting protocols from various labs into a single wiki for us to all have available | |||
** Golbeck lab now uses qwartzy for their protocol organization | |||
** Dave controls Gilmour lab’s protocols, but everyone mostly uses their own procedures | |||
** Joseph is going to set up a wiki within OpenWetWare | |||
'''Tips/Tricks''' | |||
* SLiCE cloning | |||
** Like InFusion, but is recombination based and (according to the paper) allows you to “loop out” regions during cloning | |||
** Can make your own enzyme with a very simple protocol from a bacterial strain | |||
*** Joseph obtained strain from original authors | |||
** Email paper to Brooks, Doug, Karla, Mike | |||
* Western blotting | |||
** Can use TBST (0.5% tween) for blocking during western blotting rather than milk/BSA | |||
*** Works for both nitrocellulose and PVDF | |||
** Competent cells | |||
*** Simple protocol (Inoue et al 1990) for making competent E. coli a ~109 cfu/ug efficiency | |||
*** Email paper to Brooks, Doug, Karla, Mike |
Revision as of 11:10, 27 October 2014
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Karla
- Engineer other bacteria to photosynthesize (bacillus and e coli)
- Find transformation system for Heliobacteria
- Anerobic jar’s don’t work great, little room for plates
- Stack w/stick or paper towel roll to have more plates per jar
- Heliobacter grow to slow/don’t grow, problems with temp
- Try top agar or pour agar
- Anerobic jar’s don’t work great, little room for plates
Mike
- Bacolovirus doesn’t want to express NELF, kills them?
- Try using a signal peptide to target the proteins for secretion to get them away from the cells and hopefully decrease toxicity
Cell-free expression
- New cell-free expression system with dual rxn setup (separated by membrane). Synthesis rxn and feeder rxn for substrate to diffuse from and inhibitory products to fuse into.
- Email Doug
Wiki
- Bede had previously had a good idea about collecting protocols from various labs into a single wiki for us to all have available
- Golbeck lab now uses qwartzy for their protocol organization
- Dave controls Gilmour lab’s protocols, but everyone mostly uses their own procedures
- Joseph is going to set up a wiki within OpenWetWare
Tips/Tricks
- SLiCE cloning
- Like InFusion, but is recombination based and (according to the paper) allows you to “loop out” regions during cloning
- Can make your own enzyme with a very simple protocol from a bacterial strain
- Joseph obtained strain from original authors
- Email paper to Brooks, Doug, Karla, Mike
- Western blotting
- Can use TBST (0.5% tween) for blocking during western blotting rather than milk/BSA
- Works for both nitrocellulose and PVDF
- Competent cells
- Simple protocol (Inoue et al 1990) for making competent E. coli a ~109 cfu/ug efficiency
- Email paper to Brooks, Doug, Karla, Mike
- Can use TBST (0.5% tween) for blocking during western blotting rather than milk/BSA