Team:Johns Hopkins/Notebook/GROUP 4: Long Two-way Stops & Mat(alpha) specific promoters

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.
 Gel photograph: [http://baderlab.bme.jhu.edu:8888/moodle18/mod/data/view.php?d=16&rid=4986 Alpha Promoters and Long Terminator Digest, Kan BBV]
 Gel Annotations: [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-10-29.Alpha%20Promoter%20and%20Long%20Terminator%20Digest%20in%20Kan%20BBV.Alpha%20Promoter%20and%20Long%20Terminator%20Digest%20in%20Kan%20BBV.html Alpha Promoter and Long Terminator Digest in Kan BBV]
 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.
 Gel photo: [http://baderlab.bme.jhu.edu:8888/moodle18/mod/data/view.php?d=16&rid=4987 Alpha Promoters and Long Terminators, Digestion from pGEM-T]
 Gel annotations: [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-10-29.Alpha%20Promoter%20and%20Long%20Terminator%20Digest,%20pGEM-T%20vectors.Alpha%20Promoter%20and%20Long%20Terminator%20Digest,%20pGEM-T%20vectors.html Alpha Promoter and Long Terminator Digest, pGEM-T vectors]
 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.