Endy:BAC Subgroup: Difference between revisions
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*This week | *This week | ||
==Week of 7/ | ==Week of 7/17/17== | ||
'''Alec''' | '''Alec''' | ||
Last week - | Last week - Tested the prototype of the program and generated multiple (10) primers for PURE parts. Tested the PCR once and all failed; will retry to see if failure is due to primer design or experimental setup. Also made a list of reactions that occur in PURE in preparation to build the PURE model. | ||
This week - Re-PCR the 5 components that I have designed via the program and see if amplification occurs. Build the model of PURE based on the Katy Wei thesis to be presentable at the Build-A-Cell meetup in Pasadena. | |||
'''Anton''' | '''Anton''' | ||
Line 22: | Line 23: | ||
'''Cynthia''' | '''Cynthia''' | ||
*''Reconstitution of E. coli RNA polymerase in PURE'' - | *''Reconstitution of E. coli RNA polymerase in PURE'' - Waiting on sequencing primers for two more subunits to come in, will do transformations this week. | ||
*''Characterization of E. coli RNA polymerase promoter library'' - E. coli | *''Characterization of E. coli RNA polymerase promoter library'' - Plasmid for positive control was cloned by Eric over the weekend and all promoters were PCR amplified, will do experiments with purchased E. coli RNA polymerase this week, starting with J23119 promoter. | ||
*''Liposome formation'' - | *''Liposome formation'' - Planning liposome experiments with Texas Red and egg PC | ||
'''Eric''' | '''Eric''' | ||
*'' | *''Make corrections to initiator tRNA structure and order ultramers'' - Reading old tRNA papers to figure out potential failure mode for nonfunctional initiator tRNA | ||
*''Purification of fmt enzyme'' - | *''Purification of fmt enzyme'' - Underwent purification, will run on gel to check if successful | ||
* | * Clone and purify out 6 more PURE proteins | ||
* | * Read into potential maturation enzymes necessary for the maturation of the initiator tRNA | ||
Line 50: | Line 51: | ||
'''Vicky''' | '''Vicky''' | ||
*SUMMER PROJECT: | *SUMMER PROJECT: reconstructing the 50S subunit of the ribosome. This includes synthesizing its different parts, expressing them in PURE, and coupling it with native 30S to text for functionality. | ||
*''Put together latest work on ribosome biogenesis'' - | *''Put together latest work on ribosome biogenesis'' - constantly looking for updates | ||
*''Synthesizing primers for ribosomal genes'' - Alec | *''Synthesizing primers for ribosomal genes'' - working with Alec to create a program that allows us to create primers specific to create our vector (would need it for over 110 genes) | ||
* | |||
*"This week": continue to purify more ribosomal components into vector and get them to express in pure + look for other labs that se an automated method of cloning as opposed to monocloning |
Revision as of 09:54, 17 July 2017
BAC Weekly Task List
List of planned work for the week for each individual on team
Suggested format:
- Last week - Progress
- This week
Week of 7/17/17
Alec
Last week - Tested the prototype of the program and generated multiple (10) primers for PURE parts. Tested the PCR once and all failed; will retry to see if failure is due to primer design or experimental setup. Also made a list of reactions that occur in PURE in preparation to build the PURE model.
This week - Re-PCR the 5 components that I have designed via the program and see if amplification occurs. Build the model of PURE based on the Katy Wei thesis to be presentable at the Build-A-Cell meetup in Pasadena.
Anton
Conary
Cynthia
- Reconstitution of E. coli RNA polymerase in PURE - Waiting on sequencing primers for two more subunits to come in, will do transformations this week.
- Characterization of E. coli RNA polymerase promoter library - Plasmid for positive control was cloned by Eric over the weekend and all promoters were PCR amplified, will do experiments with purchased E. coli RNA polymerase this week, starting with J23119 promoter.
- Liposome formation - Planning liposome experiments with Texas Red and egg PC
Eric
- Make corrections to initiator tRNA structure and order ultramers - Reading old tRNA papers to figure out potential failure mode for nonfunctional initiator tRNA
- Purification of fmt enzyme - Underwent purification, will run on gel to check if successful
- Clone and purify out 6 more PURE proteins
- Read into potential maturation enzymes necessary for the maturation of the initiator tRNA
Keoni
- Last week: Collection of materials for testing Bacillus subtilis minipreping and storage. Tested conjugation with Vibrio natriegens.
- This week: Preparing DNA synthesis sequences.
Maria
Last week:
- Successfully transformed the conjugative plasmid pRK24 into recipient strain MG1AC53.
- Next step on this confirming the presence of pRK24 with gel.
This week:
- Read literature on no-SCAR system with the purpose of replacing the oriT-kan casette with negative selection marker SacB on pRK24. Design appropriate sequences to carry out this experiment. This would be done to avoid back-conjugation of the genome from the recipient to the donor.
Vicky
- SUMMER PROJECT: reconstructing the 50S subunit of the ribosome. This includes synthesizing its different parts, expressing them in PURE, and coupling it with native 30S to text for functionality.
- Put together latest work on ribosome biogenesis - constantly looking for updates
- Synthesizing primers for ribosomal genes - working with Alec to create a program that allows us to create primers specific to create our vector (would need it for over 110 genes)
- "This week": continue to purify more ribosomal components into vector and get them to express in pure + look for other labs that se an automated method of cloning as opposed to monocloning