IGEM:MIT/2006/Notebook/2006-8-31: Difference between revisions

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==to do==
==to do==
#miniprep iGEM yeast shuttle vector and digest it appropriately (check the MCS, or Veena took notes and might know). Also digest our BGD (J45200,J45220) appropriately to insert it at the MCS
#dilute 4 plate reader LCs 1:500 and load w/Barry at 9 am '''-done'''
#analyze incoming sequencing extra well b/c are final bb-box constructs (and throw out duplicates if these all look good)
#glycerol (6) J45399 LCs (note: 2XYT problem) '''-done''' and miniprep '''-done'''
#primers should be arriving today, so we should try to PCR pmsCEAB-full out of PE3 and pE3R (minipreps of both plasmids from 8/29) '''-done/in progress'''
#check if the 0-Q-199A forward sequencing is in from the re-run '''-done''' (it's all good)
#sequence (6) J45399 attempts (b/c similar # of colonies on 2nd attempt I think its worth a try) '''-done'''
#miniprep iGEM yeast shuttle vector '''-done'''
#digest yeast with XS. Also digest our ATF1 biobrick (J45014) with XS. '''NOTE:''' we need to do freeze step so that X and S don't religate for both of these!!
#ligate ATF1 into yeast shuttle vector and transform into yeast (all in one step, hopefully samantha is around)
#pellet big yeast LC with big centrifuge and resuspend cells in milk and find a good bread recipe for baking
#pick colonies from all 8/30 transformant plates and make LCs
#check on pseudomonas '''-done'''
 
==to do tomorrow==
#analyze the incoming 12 sample sequencing order (and throw out duplicates/bad ones) '''done, also made new LCs for failed sequences'''
 
==also==
#'''UPDATE REGISTRY''': create registry biobrick parts and choose numbers for our inverter constructs (w/ and w/out osmY attached) '''as well as put all intermediate BGD, IAGD, SAGD parts into the registry!!!!''' - p.s. anybody out of town is welcome to help. we seem to be very behind on this.

Latest revision as of 15:26, 31 August 2006

to do

  1. dilute 4 plate reader LCs 1:500 and load w/Barry at 9 am -done
  2. glycerol (6) J45399 LCs (note: 2XYT problem) -done and miniprep -done
  3. primers should be arriving today, so we should try to PCR pmsCEAB-full out of PE3 and pE3R (minipreps of both plasmids from 8/29) -done/in progress
  4. check if the 0-Q-199A forward sequencing is in from the re-run -done (it's all good)
  5. sequence (6) J45399 attempts (b/c similar # of colonies on 2nd attempt I think its worth a try) -done
  6. miniprep iGEM yeast shuttle vector -done
  7. digest yeast with XS. Also digest our ATF1 biobrick (J45014) with XS. NOTE: we need to do freeze step so that X and S don't religate for both of these!!
  8. ligate ATF1 into yeast shuttle vector and transform into yeast (all in one step, hopefully samantha is around)
  9. pellet big yeast LC with big centrifuge and resuspend cells in milk and find a good bread recipe for baking
  10. pick colonies from all 8/30 transformant plates and make LCs
  11. check on pseudomonas -done

to do tomorrow

  1. analyze the incoming 12 sample sequencing order (and throw out duplicates/bad ones) done, also made new LCs for failed sequences

also

  1. UPDATE REGISTRY: create registry biobrick parts and choose numbers for our inverter constructs (w/ and w/out osmY attached) as well as put all intermediate BGD, IAGD, SAGD parts into the registry!!!! - p.s. anybody out of town is welcome to help. we seem to be very behind on this.
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