IGEM:MIT/2007/Notebook/2007-8-7

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(New page: <pre> 8-7 Minutes update Talked about polystyrene binding-works Mercury binding-ordered mercury sensing-not working compare random sequence to e coli genome then make part from it talke...)
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Things we need to do are starred
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8-7
8-7

Revision as of 13:50, 7 August 2007

Things we need to do are starred

8-7
Minutes

update
Talked about polystyrene binding-works
Mercury binding-ordered
mercury sensing-not working

compare random sequence to e coli genome
then make part from it

talked about polystyrene assay 
*need to do negative control of (no plasmid)
*check specificity and try polypropylene etc
*arrange in order of increasing concentrations
*do viable count of bacteria at high concentrations

t7 fluorescent antibody tag
*scale bar
*bright+fluorescent
Xis it same scale across all images or does program chooses intensity. eric answered this
*control of positive control antibody and negative of antibody to demonstrate antibody bound.  
*do a secondary antibody only test
*hi res of stained antibody-can we tell localized binding or to polystyrene?


what's next
we've done in pbs, but need to do in water
need to put cpx downstream of promoter

metal binding
using ec20 on lpp-ompa
*find out how well iron binds to ec20
*find out toxic levels for similar metals (iron, zinc).  do background check on this


Mike Bennie:<<ask him
biobricking beta c term domain of n terminus
~1 week away; most pieces are done.

MTA with de Lorenzo.
protein export tag
fos gune??
leucine zipper
beta portion of 84
ege1 has protein autotransporter



*Possible leads on psb3k3 purifications:
*let tk know; can watch
*do chlorampheticol amplification on plasmid
grow overnight, dilute, regrow in chlorampheticol; inhibits protein production but not dna replication->all of energy goes into copying; 10x yield on low copy plasmids. (must check if not chloramphenicol resistant)
*use TE instead of water
*can heat it; if heat elution buffer to 50 or 60 degrees,
*elute by 30 ul then combine 30 ul


*what percentage of bacteria is binding?
*try fine particulate polystyrene


**engineering fish to prevent mercury acculumation

**nitrocellulose sandwiches
*robbie's a pro at this stuff
robbie or rana for western blot

thurs 10 am 4-261
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