Team:Johns Hopkins/Notebook/GROUP 2: MATa Specific-promoters

GROUP 2: MATa Specific-promoters

 Status report by Allison and Nate
 Part no.: BBa_K110008
 Part Description: MFA1 (L+R)
 Part Location: in a labeled box, second shelf from the top, -20
 degrees C refrigerator next to front door
 Date: 7/10/08
 PCR successful? Yes
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.BioBricks%208%20and%2016.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html BioBricks 8 and 16]
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.BioBricks%208%20and%2016%20Using%20Constant%20Annealing.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html BioBricks 8 and 16 Using Constant Annealing]
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.Short%202Way%20Stop,%20Alpha%20Promoters,%20&%20Sapphire%20FP.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html Short 2Way Stop, Alpha Promoters, & Sapphire FP]
 Cloning of PCR product successful: in progress
 Sequencing of cloned PCR product successful: not done
 Joining of validated part to adjacent part(s) status: not done
 Problems to be solved: to be determined
 Current status of this part: PCR was being troubleshooted, appeared to
 have good results with regular PCR protocol (not touchdown) in which
 there was a constant annealing temperature of 55 degrees C - see gel

 Status report by Allison and Nate
 Part no.: BBa_K110016
 Part Description: Ste2 (R+L)
 Part Location: in a labeled box, second shelf from the top, -20
 degrees C refrigerator next to front door
 Date: 7/10/08
 PCR successful? Yes
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.BioBricks%208%20and%2016.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html BioBricks 8 and 16]
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.BioBricks%208%20and%2016%20Using%20Constant%20Annealing.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html BioBricks 8 and 16 Using Constant Annealing]
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.Short%202Way%20Stop,%20Alpha%20Promoters,%20&%20Sapphire%20FP.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html Short 2Way Stop, Alpha Promoters, & Sapphire FP]
 Cloning of PCR product successful: in progress
 Sequencing of cloned PCR product successful: not done
 Joining of validated part to adjacent part(s) status: not done
 Problems to be solved: to be determined
 Current status of this part: Both PCR protocols (touchdown and second
 PCR with constant annealing temperature) produced product of the
 correct size. BBa_K110016 was used as a control in the second PCR with
 BBa_K110008.

 Status report by Allison and Nate
 Part no.: BBa_K110008
 Part Description: MFA1 (L+R)
 Part Location: same as above, plates are at 4 degrees refrigerator near front door
 Date: 7/14/08
 PCR successful? Yes
 Cloning of PCR product of successful? There were mainly light blue colonies 
  (only a couple white colonies)
 Sequencing of cloned PCR product successful: not done
 Joining of validated part to adjacent part(s) status: not done
 Problems to be solved: Ligation
 Current status of this part: plates are at 4 degrees; another ligation/transformation 
  will be completed soon

 Status report by Allison and Nate
 Part no.: BBa_K110016
 Part Description: Ste2 (R+L)
 Part Location: in a labeled box, second shelf from the top, -20 degrees C refrigerator next to 
  front door; plates at 4 degrees
 Date: 7/14/08
 PCR successful? Yes
 Cloning of PCR product successful: There were many blue colonies (similar to the plate of BB_K110008)
 Sequencing of cloned PCR product successful: not done
 Joining of validated part to adjacent part(s) status: not done
 Problems to be solved: Ligation
 Current status of this part: plates are at 4 degrees; another ligation/transformation 
  will be completed soon

 Status report by Allison and Nate
 Part no.: BBa_K110008
 Part Description: MFA1 (L+R)
 Part Location: same as above, plates are at 4 degrees refrigerator near front door
 Date: 7/22/08
 PCR successful? Yes 
 (BAD LINK)
 Cloning of PCR product of successful? Yes (approx 60 white colonies between two plates)
 Sequencing of cloned PCR product successful: not done
 Joining of validated part to adjacent part(s) status: not done
 Problems to be solved:
 Current status of this part: 12 mini preps and the restriction digestion were completed;
 preparing to send 3 samples to be sequenced
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.Restriction%20Digests%20of%20BBa_K1100...%2008%20&%2016.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html Restriction Digests of BBa_K1100... 08 & 16]

 Status report by Allison and Nate
 Part no.: BBa_K110016
 Part Description: Ste2 (R+L)
 Part Location: in a labeled box, second shelf from the top, -20 degrees C refrigerator next to 
  front door; plates at 4 degrees
 Date: 7/22/08
 PCR successful? Yes
 Cloning of PCR product successful: Yes (approx 20 white colonies on one plate)
 Joining of validated part to adjacent part(s) status: not done
 Problems to be solved:
 Current status of this part: 12 mini preps and the restriction digestion were completed;
 preparing to send 3 samples to be sequenced
 [http://www.jhu.edu/iGEM/Group2:MATaSpecificPromoters/2008-7-22.Restriction%20Digests%20of%20BBa_K1100...%2008%20&%2016.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html Restriction Digests of BBa_K1100... 08 & 16]

 Date: July 29, 2008
 Status report by: Allison and Nate
 Part no.: BBa_K110008 -> BBa_K110016
 Part Description: A promoters: MFA1 (L+R) and Ste2 (R+L), respectively.
 Two samples of mini preps from MFA1 and Ste2 were sent off for sequencing at the end of last week.
 Results will follow soon.