Team:Johns Hopkins/Notebook/GROUP 5: MATa Specific Promoters II

GROUP 5: MATa Specific Promoters II

Summary for MATa Specific Promoters II Group

 Part Description: Ste2 Promoter Reverse
 Summary here: We were able to grow colonies with sequence verified Ste2 promoters. 
   The promoter is currently located in a glycerol stock in pGEM vector. One attempt to 
   transfer to the iGEM vector has been made, unsuccessfully. This was due to a mistake 
   in protocol however, so final transfer to the iGEM vector should still be straightforward. 
   Progress on this bio-brick has been paused in order to focus on MFA1, which is more 
   applicable to our project design because this Ste2 designed for the wrong direction.
 
 Part no.: BBa_K110015
 Part Description: MFA1 Promoter Forward
 Summary here: We have been unable to get successful, sequence verified clones with 
   our MFA1 promoter thus far. We currently have a few samples of DNA ready to be tested 
   by restriction enzyme digest, and then, hopefully, sequencing.

For more information about the parts go to: http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008& group=Johns_Hopkins

Up to today...
 Since then -- we have been troubleshooting and working on getting higher concentrations and using
 the correct stock plates. Several delays have occurred,and we should be close to ligating into the
 iGEM vector

 8/24 
 Restriction Enzyme Digested and the insert appers promising. Ready for sequence submission for Ste 2 promotor. 
 Status Report by: Alexandra McMillan

 8/21 
 Due to low concentrations, we needed to concentrate our DNA before preceeding. 
 Ethanol precipitation using EtOH/NaAc and 100 ul of miniprep mix was complete of the BBa_K110015 
 and BBa_K11009 minipreps.

 8/18
 Miniprep products were < 100 ng/ul. Restriction digest. There appear to be correct
 size inserts. Gel image coming. Results look promising.

 8/15
 DNA miniprep of BBa_K110015 and BBa_K11009 cultures

 8/13
 Incubated inoculations of old colony plates

 By 7/19/08
 We successfuly transformed both BB 15 and 9 and ran a gel that came out with faint bands. 
 We digested BB 9 and 15.

 Date: 7/11/08
 status report by Rick Carrick
 Part no.: BBa_K110015,
 Part Description: MFA1
 Part Location (in build a genome lab):
 PCR successful?; Y (on moodle somewhere)
 Cloning of PCR product successful: Y
 Sequencing of cloned PCR product successful:N
 Joining of validated part to adjacent part(s) status: Not done
 Problems to be solved: None so far
 Current status of this part: This parts must be restriction enzyme digested and sequenced 
 next.

 Date: 7/11/08
 status report by Rick Carrick
 Part no.: BBa_K110009
 Part Description: Ste2
 Part Location (in build a genome lab):
 PCR successful?; Y (on moodle somewhere)
 Cloning of PCR product successful: Y
 Sequencing of cloned PCR product successful:N
 Joining of validated part to adjacent part(s) status: Not done
 Problems to be solved: None so far
 Current status of this part: This part must be restriction enzyme digested and sequenced
 next