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: Alpha Promoters and Long Terminator Digest, Kan BBV Gel Annotations: 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: Alpha Promoters and Long Terminators, Digestion from pGEM-T Gel annotations: 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 Restriction Enzyme Digest of Mini-Preps Summary here: Need to be sequenced.