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

From 2008.igem.org

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: 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
 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
 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
 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
 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
 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