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Team:Johns Hopkins/Notebook
From 2008.igem.org
Notebook
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iGEM Groups V1.0
iGEM Groups V2.01
Important reminders and notes
[Can make general comments here, so they don't get lost in peoples e-mail boxes]
July-11: Primers for group 1 were delivered yesterday
July 11: Lab meeting at 7:30PM in the lab to go over miniprep protocol
Tuesday July 15: Lab meeting at 6:30PM with Jessica. Have status reports ready.
Status Reports
GROUP 1: Fluorescent Proteins
status report by: Ingrid (work done by James) Part no.: BBa_K110017 Part Description: yESapphire RtL Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; Yes Cloning of PCR product successful: Y/N Sequencing of cloned PCR product successful: Not done Joining of validated part to adjacent part(s) status: Not done Problems to be solved: The PCR of this part yielded a very large product Current status of this part:
status report by: Ingrid (work done by James) Part no.: BBa_K110010 Part Description: yESapphire LtR Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; Yes Cloning of PCR product successful: Not done Sequencing of cloned PCR product successful: No Joining of validated part to adjacent part(s) status: Not done Problems to be solved: The PCR of this part yielded a very large product Current status of this part:
GROUP 2: MATa Specific-promoters
Status report by Allison and Nate Part no.: BBa_K110008 Part Description: MFA1 (L+R) Part Location: in a labeled box, second shelf from the top, -20 degrees C refrigerator next to front door Date: 7/10/08 PCR successful? Yes Cloning of PCR product successful: in progress Sequencing of cloned PCR product successful: not done Joining of validated part to adjacent part(s) status: not done Problems to be solved: to be determined Current status of this part: PCR was being troubleshooted, appeared to have good results with regular PCR protocol (not touchdown) in which there was a constant annealing temperature of 55 degrees C - see gel
Status report by Allison and Nate Part no.: BBa_K110016 Part Description: Ste2 (R+L) Part Location: in a labeled box, second shelf from the top, -20 degrees C refrigerator next to front door Date: 7/10/08 PCR successful? Yes Cloning of PCR product successful: in progress Sequencing of cloned PCR product successful: not done Joining of validated part to adjacent part(s) status: not done Problems to be solved: to be determined Current status of this part: Both PCR protocols (touchdown and second PCR with constant annealing temperature) produced product of the correct size. BBa_K110016 was used as a control in the second PCR with BBa_K110008.
GROUP 3: Short two way stops
Date: 7/11/08 status report by: James Part no.: BBa_K110011 Part Description: Between-bud 27-W FRS2-C LtR Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; Yes Cloning of PCR product successful: Yes (will come soon; I can put it in the wiki to make it easier for you) Sequencing of cloned PCR product successful: No Joining of validated part to adjacent part(s) status: Not done Problems to be solved: Current status of this part: Miniprep of Overnight cultures will be completed today
status report by: James Part no.: BBa_K110012 Part Description: Between STE2-W and BST1-C LtR Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; Yes Cloning of PCR product successful: Yes (will come soon; I can put it in the wiki to make it easier for you) Sequencing of cloned PCR product successful: No Joining of validated part to adjacent part(s) status: Not done Problems to be solved: Current status of this part: Miniprep of Overnight cultures will be completed today
status report by: James Part no.: BBa_K110013 Part Description: Between-SWP82-W and EMP47-C LtR Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; No Cloning of PCR product successful: No (will come soon; I can put it in the wiki to make it easier for you) Sequencing of cloned PCR product successful: No Joining of validated part to adjacent part(s) status: Not done Problems to be solved: The PCR of this part yielded a very large product Current status of this part:
GROUP 4: Long Two-way Stops & Mat(alpha) specific promotors
[Jaime- Please split]
Date: 7/10/08 Status report by: Jaime Liu Part no.: BBa_K110001, BBa_K110003, BBa_K110005, BBa_K110006 Part Description: BBa_K110001 - Between-bud 27-W FRS2-C + 200bp into each gene LtR BBa_K110003 - Between-SWP82-W and EMP47-C +200 into each gene LtR BBa_K110005 - MFalpha2 LtR BBa_K110006 - MFalpha1 LtR Part Location (in build a genome lab): In 4C fridge #2 PCR successful?; Y/N (link such as this)- Yes BBa_K110001, BBa_K110003: http://baderlab.bme.jhu.edu:8888/moodle18/mod/data/view.php?d=4&rid=1462 http://baderlab.bme.jhu.edu:8888/moodle18/mod/data/view.php?d=4&rid=1463 BBa_K110005, BBa_K110006: http://baderlab.bme.jhu.edu:8888/moodle18/mod/data/view.php?d=4&rid=1470 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 1.5 mL LB for mini-prep.
GROUP 5: MATa Specific Promoters II
Date: 7/11/08 status report by Rick Carrick Part no.: BBa_K110015, Part Description: MFA1 Part Location (in build a genome lab): PCR successful?; Y (on moodle somewhere) Cloning of PCR product successful: Y Sequencing of cloned PCR product successful:N Joining of validated part to adjacent part(s) status: Not done Problems to be solved: None so far Current status of this part: This parts must be restriction enzyme digested and sequenced next.
Date: 7/11/08 status report by Rick Carrick Part no.: BBa_K110009 Part Description: Ste2 Part Location (in build a genome lab): PCR successful?; Y (on moodle somewhere) Cloning of PCR product successful: Y Sequencing of cloned PCR product successful:N Joining of validated part to adjacent part(s) status: Not done Problems to be solved: None so far Current status of this part: This part must be restriction enzyme digested and sequenced next
GROUP 6: Vectors
Status report by ____ Vector transformed into bacteria strain DB3.1 Y/N Permanent culture made in the Boeke lab for future reference Y/N Selectable marker for this vector Medium made and tested Y/N (link) DNA preps made Y/N DNA preps tested by RE digest - (link) DNA preps tested by transformation into DB3.1 and DH5alpha (or JM109) – put data as Table on moodle. Sample format/data follows:
Amount transformed cfu/micGm in DB3.1 cfu/micGm in JM109 0.1 ng 5 * 10e7 <2 * 10e2
Preparative digests ready for use are located – where?
GROUP 7
