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Team:Johns Hopkins/Notebook/GROUP 2: MATa Specific-promoters
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<b>Two samples of mini preps from MFA1 and Ste2 were sent off for sequencing at the end of last week. | <b>Two samples of mini preps from MFA1 and Ste2 were sent off for sequencing at the end of last week. | ||
Results will follow soon.</b> | Results will follow soon.</b> | ||
| + | |||
| + | Date: August 5, 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. | ||
| + | <b>Sequences were analyzed. BBa_K110016 had a perfect clone. Since there is not much | ||
| + | miniprep left, a transformation will be done to generate more clones with the | ||
| + | correct sequence. BBa_K110008 had one mutation; additional minipreps from other clones | ||
| + | are ready to send off for sequencing, although the DNA concentrations are low. Colonies | ||
| + | can be picked and overnight cultures grown | ||
| + | for another round of minipreps.</b> | ||
Revision as of 01:26, 13 August 2008
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.
Date: August 5, 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. Sequences were analyzed. BBa_K110016 had a perfect clone. Since there is not much miniprep left, a transformation will be done to generate more clones with the correct sequence. BBa_K110008 had one mutation; additional minipreps from other clones are ready to send off for sequencing, although the DNA concentrations are low. Colonies can be picked and overnight cultures grown for another round of minipreps.
