Team:Johns Hopkins/Notebook/GROUP 4: Long Two-way Stops & Mat(alpha) specific promoters
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- | == GROUP 4: Long Two-Way Stops & MAT(alpha) Specific Promoters== | + | |
+ | == GROUP 4: Long Two-Way Stops & MAT(alpha) Specific Promoters == | ||
+ | |||
+ | <b>Summary for Long Two-Way Stops & MAT(alpha) Specific Promoters Group</b> | ||
+ | |||
+ | 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] | ||
+ | |||
+ | Summary here: | ||
+ | Need to be sequenced. |
Revision as of 01:18, 30 October 2008
GROUP 4: Long Two-Way Stops & MAT(alpha) Specific Promoters
Summary for Long Two-Way Stops & MAT(alpha) Specific Promoters Group
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] Summary here: Need to be sequenced.