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Team:Johns Hopkins/Notebook/GROUP 4: Long Two-way Stops & Mat(alpha) specific promotors
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| - | == GROUP 4: Long Two-way Stops & Mat(alpha) specific | + | == GROUP 4: Long Two-way Stops & Mat(alpha) specific promoters == |
| + | |||
| + | Summary: As of September 23rd, 2008 | ||
| + | We had successfully cloned BBa_K110005 and BBa_K110006 (alpha Promoters) and | ||
| + | BBa_K110003 (Long Terminator) into pGEM-T and Kanamycin biobrick vectors. | ||
| + | They await assembly and ligation. | ||
| + | |||
| + | Date: September 17th, 2008 | ||
| + | Minipreps were made of the kanamycin biobrick vector clones, and digested with E/P to | ||
| + | check for insert of correct size. See gel: /pic// | ||
| + | Low-copy origin makes for poor gel visualization. Suggest some other diagnostic test. | ||
| + | |||
| + | Date: September 12th, 2008 | ||
| + | Clones were digested with E/P, and ligated O/N into Kanamycin biobrick vector (pSB4K5). | ||
| + | Colonies were picked and grown. | ||
| + | |||
| + | Date: August 26th, 2008 | ||
| + | Author: Jonathan | ||
| + | Sequences were returned, and prefix/suffix/identities were confirmed. Clones that passed are: | ||
| + | Long Terminator BBa_K110003: Clone A1-3.2 | ||
| + | Alpha Promoter BBa_K110005: Clone 5.1 | ||
| + | Alpha Promoter BBa_K110006: Clones 6.1, 6.3 | ||
| + | |||
| + | Date: August 21st, 2008 | ||
| + | Author: Jonathan | ||
| + | Clones were grown up O/N and re-miniprepped. Digestion with E/P yielded some possible candidates. | ||
| + | Here is the gel pic: [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-10-29.08/21/08%20E/P%20Restriction%20Digest%20for%20BBs%203,%205,%206%20(pGEM-T).Alpha%20Promoters/Long%20Terminators%20Restriction%20Digest.html 08/21/08 E/P Restriction Digest for BBs 3, 5, 6 (pGEM-T)] | ||
| + | Clones submitted for sequencing. | ||
| + | |||
| + | Date: August 1st, 2008 | ||
| + | Author: Jonathan | ||
| + | Sequences were submitted to the facility (3/building block) and a few returned perfect clones, but | ||
| + | due to miscommunication, exactly which clones that the trace files correspond to are under some | ||
| + | confusion. Will attempt to confirm clone identities. | ||
| + | |||
| + | Date: July 29, 2008 | ||
| + | Status report by: Jaime | ||
| + | Part no.: BBa_K110001, BBa_K110003, BBa_K110005, BBa_K110006 | ||
| + | Part Description: Long Two-way Stops & Mat(alpha) specific promotors | ||
| + | [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-25.Restriction%20Enzyme%20Digest%20of%20Mini-Preps.Jaime.html Restriction Enzyme Digest of Mini-Preps] | ||
| + | |||
| + | |||
Date: 7/10/08 | Date: 7/10/08 | ||
| Line 25: | Line 66: | ||
Part Location (in build a genome lab): In 4C fridge #2 | Part Location (in build a genome lab): In 4C fridge #2 | ||
PCR successful?; Y/N (link such as this)- Yes | PCR successful?; Y/N (link such as this)- Yes | ||
| - | + | [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-22.iGEM%20PCR%20experimentation%202%20on%20BBa_K110001%20and%20BBa_K110003.Jaime.html iGEM PCR experimentation 2 on BBa_K110001 and BBa_K110003] | |
Cloning of PCR product successful: Y/N Yes | Cloning of PCR product successful: Y/N Yes | ||
Sequencing of cloned PCR product successful: Y/N No | Sequencing of cloned PCR product successful: Y/N No | ||
| Line 40: | Line 81: | ||
Part Location (in build a genome lab): In 4C fridge #2 | Part Location (in build a genome lab): In 4C fridge #2 | ||
PCR successful?; Y/N (link such as this)- Yes | PCR successful?; Y/N (link such as this)- Yes | ||
| - | + | [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-22.iGEM%20BBa_K110005%20and%20BBa_K110006.Jaime.html iGEM BBa_K110005 and BBa_K110006] | |
Cloning of PCR product successful: Y/N Yes | Cloning of PCR product successful: Y/N Yes | ||
Sequencing of cloned PCR product successful: Y/N No | Sequencing of cloned PCR product successful: Y/N No | ||
| Line 55: | Line 96: | ||
Part Location (in build a genome lab): In 4C fridge #2 | Part Location (in build a genome lab): In 4C fridge #2 | ||
PCR successful?; Y/N (link such as this)- Yes | PCR successful?; Y/N (link such as this)- Yes | ||
| - | + | [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-22.iGEM%20BBa_K110005%20and%20BBa_K110006.Jaime.html iGEM BBa_K110005 and BBa_K110006] | |
Cloning of PCR product successful: Y/N Yes | Cloning of PCR product successful: Y/N Yes | ||
Sequencing of cloned PCR product successful: Y/N No | Sequencing of cloned PCR product successful: Y/N No | ||
Latest revision as of 00:58, 30 October 2008
GROUP 4: Long Two-way Stops & Mat(alpha) specific promoters
Summary: As of September 23rd, 2008 We had successfully cloned BBa_K110005 and BBa_K110006 (alpha Promoters) and BBa_K110003 (Long Terminator) into pGEM-T and Kanamycin biobrick vectors. They await assembly and ligation.
Date: September 17th, 2008 Minipreps were made of the kanamycin biobrick vector clones, and digested with E/P to check for insert of correct size. See gel: /pic// Low-copy origin makes for poor gel visualization. Suggest some other diagnostic test. Date: September 12th, 2008 Clones were digested with E/P, and ligated O/N into Kanamycin biobrick vector (pSB4K5). Colonies were picked and grown.
Date: August 26th, 2008 Author: Jonathan Sequences were returned, and prefix/suffix/identities were confirmed. Clones that passed are: Long Terminator BBa_K110003: Clone A1-3.2 Alpha Promoter BBa_K110005: Clone 5.1 Alpha Promoter BBa_K110006: Clones 6.1, 6.3
Date: August 21st, 2008 Author: Jonathan Clones were grown up O/N and re-miniprepped. Digestion with E/P yielded some possible candidates. Here is the gel pic: [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-10-29.08/21/08%20E/P%20Restriction%20Digest%20for%20BBs%203,%205,%206%20(pGEM-T).Alpha%20Promoters/Long%20Terminators%20Restriction%20Digest.html 08/21/08 E/P Restriction Digest for BBs 3, 5, 6 (pGEM-T)] Clones submitted for sequencing.
Date: August 1st, 2008 Author: Jonathan Sequences were submitted to the facility (3/building block) and a few returned perfect clones, but due to miscommunication, exactly which clones that the trace files correspond to are under some confusion. Will attempt to confirm clone identities.
Date: July 29, 2008 Status report by: Jaime Part no.: BBa_K110001, BBa_K110003, BBa_K110005, BBa_K110006 Part Description: Long Two-way Stops & Mat(alpha) specific promotors [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-25.Restriction%20Enzyme%20Digest%20of%20Mini-Preps.Jaime.html Restriction Enzyme Digest of Mini-Preps]
Date: 7/10/08 Status report by: Jaime Liu Part no.: BBa_K110001 Part Description: BBa_K110001 - Between-bud 27-W FRS2-C + 200bp into each gene LtR Part Location (in build a genome lab): In 4C fridge #2 PCR successful?; Y/N (link such as this)- Yes [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-22.iGEM%20PCR%20experimentation%20on%20BBa_K110001%20and%20BBa_K110003.Jaime.html iGEM PCR experimentation on BBa_K110001 and BBa_K110003] Cloning of PCR product successful: Y/N Yes Sequencing of cloned PCR product successful: Y/N No Joining of validated part to adjacent part(s) status: Not Done Problems to be solved: Not really a problem, but need do a mini-Prep and sequence Current status of this part: All cloned and inoculated into 100uL in 96 well plate for sequencing- put in incubator on 7/15
Date: 7/10/08 Status report by: Jaime Liu Part no.: BBa_K110003 Part Description: BBa_K110003 - Between-SWP82-W and EMP47-C +200 into each gene LtR Part Location (in build a genome lab): In 4C fridge #2 PCR successful?; Y/N (link such as this)- Yes [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-22.iGEM%20PCR%20experimentation%202%20on%20BBa_K110001%20and%20BBa_K110003.Jaime.html iGEM PCR experimentation 2 on BBa_K110001 and BBa_K110003] Cloning of PCR product successful: Y/N Yes Sequencing of cloned PCR product successful: Y/N No Joining of validated part to adjacent part(s) status: Not Done Problems to be solved: Not really a problem, but need do a mini-Prep and sequence Current status of this part: All cloned and inoculated into 100uL in 96 well plate for sequencing- put in incubator on 7/15
Date: 7/10/08 Status report by: Jaime Liu Part no.: BBa_K110005 Part Description: BBa_K110005 - MFalpha2 LtR Part Location (in build a genome lab): In 4C fridge #2 PCR successful?; Y/N (link such as this)- Yes [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-22.iGEM%20BBa_K110005%20and%20BBa_K110006.Jaime.html iGEM BBa_K110005 and BBa_K110006] Cloning of PCR product successful: Y/N Yes Sequencing of cloned PCR product successful: Y/N No Joining of validated part to adjacent part(s) status: Not Done Problems to be solved: Not really a problem, but need do a mini-Prep and sequence Current status of this part: All cloned and inoculated into 100uL in 96 well plate for sequencing- put in incubator on 7/15
Date: 7/10/08 Status report by: Jaime Liu Part no.: BBa_K110006 Part Description: BBa_K110006 - MFalpha1 LtR Part Location (in build a genome lab): In 4C fridge #2 PCR successful?; Y/N (link such as this)- Yes [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-22.iGEM%20BBa_K110005%20and%20BBa_K110006.Jaime.html iGEM BBa_K110005 and BBa_K110006] Cloning of PCR product successful: Y/N Yes Sequencing of cloned PCR product successful: Y/N No Joining of validated part to adjacent part(s) status: Not Done Problems to be solved: Not really a problem, but need do a mini-Prep and sequence Current status of this part: All cloned and inoculated into 100uL in 96 well plate for sequencing- put in incubator on 7/15
