===== Minutes ===== ====January 2012==== __Monday, 30th__ - Came up with a weekly meeting schedule (Tuesday, 2-4 p.m.) - Discussed the fundamental biological concepts behind the quantum dots - Dionne and Allison discussed the background of the iGem competition - Also gave an overview of the project's direction __Tuesday, 31st__ -Anthony, Eric and Yuta attended training with Dionne. -Learned fundamentals of microbiology -Laboratory tour and practical -Pipette Skills Test *Eric scored 0.996g *Yuta scored 0.989g *Anthony scored 0.986g ====February 2012==== __Wednesday, 1st__ - Met and discussed each of the eight individual pages of sketches - Decided on the six ideas to present to the class - Drew up a preliminary problem statement __Tuesday, 7th__ - Review preliminary ideas presented in last Friday's EID crit - Assigned to calculate basic molar concentration/create MSDS - Learned to use partsregistry - Developed technique in assessing required parts for bacterial plasmid - Scheduled this week's training sessions for Friday (will come in pairs) __Friday, 10th__ - Lab safety tutorial - DNA introduction - Electrophoresis theory/practical - Hand washing "test" *Anthony passed *Stanley passed *Yuta failed *Eric failed __Tuesday, 14th__ - Reviewed individual research of past iGEM projects on possible group project - Learned the basics of bacteria, DNA, and basic transformation protocol - Produced a basic timeline for project - Scheduled additional training sessions for Wednesday (will come in pairs) __Wednesday, 15th__ - Bacterial transformation theory/practical - Reviewed procedure of transforming bacterial DNA __Tuesday, 21st__ - Theory on restriction enzymes - Review the basic protocol of using restriction enzymes for bacterial transformation - Practical for making the gel for the gel electrophoresis (gel made at different concentrations) - Practical for bacterial streaking - Plan for re-experimentation of last summer's iGEM team's work-date projected: next Friday - Hand washing "test" *Anthony passed *Stanley passed *Yuta passed *Eric passed __Tuesday, 28th__ - Plan for experiment on Friday - Review and produce protocol to synthesize CdS (QD) - Discuss possible modifications to experiment-may conduct some of these on Friday ====March 2012==== __Friday, 2nd__ - Review and make last minute modification to protocol - Confirm calculation for CdCl2 and Na2S⋅9H2O to be measured during the experiment - Execute experimental protocol - Please check the lab work section for exact procedure and lab photos - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] - Go to [[start:classes:principlesofdesign:igem:photographs|Lab Photos]] __Wednesday, 14th (pi day)__ - Review past SDS page design - Draw rough sketch of our own "homemade" SDS page (based on the given restriction-108x108mm gel) - Develop elementary measurements for the SDS page - Design a prototype on SolidWorks - Create a drawing document on SolidWorks for possible laser cutting - Discuss the safety features/electric components of SDS page - Redeveloping of design for tomorrow __Thursday, 15th__ - Identify problems with initial Solid Works model - Receive advisement for the cap portion and stability issues of SDS page with lab technician - Review design with professor/will refine design details __Friday, 16th__ - Refine and finalize the design to SDS page - Refine the "glass" to a double pane instead of single - Receive approval from professor and laser cutter technician - Need the dimensions for the electrodes - Will build in the upcoming week or two __Tuesday, 20th__ - Redevelop the problems with the leaking points in design - Consider the possibility of switching models for SDS page-concluded would like to improve upon our original design - Reestablish goals - Look at past/existing models-extract the positive aspects - Decide to meet in future with smaller groups (i.e. in pairs) to increase efficiency in work - Rebuild the SDS page model for next week __Tuesday, 27th__ - Consider and redefine building two (instead of one) models for the SDS page - Develop a schedule for upcoming experiments/lab work - Review the basic protocol for the experiment: creating pET28+CDS7 plasmid - Experiment will start tomorrow __Wednesday, 28th__ - Conduct Day 1 of experiment - Last minute change to one of the plasmid backbone - Please check the lab work section for exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 3//) __Thursday, 29th__ - Conduct Day 2 of experiment - Please check the lab work section for exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 4//) ====April 2012==== __Tuesday, 3rd__ - Conduct Day 3 of experiment - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 5//) __Wednesday, 4th__ - Conduct Day 4 of experiment - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 6//) __Thursday, 5th__ - Conduct Day 5 of experiment - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 7//) - Also note that this day's experiment was conducted in two parts __Friday, 6th__ - Conduct Day 6 of experiment - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 8//) __Monday, 9th__ - Conduct Day 7 of experiment - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 9//) - Add holes for electrodes in SolidWorks assembly of SDS page - Define the holes so the hole on case align with hole on the SDS page (vertical gel electrophoresis) __Tuesday, 10th__ - Conduct Day 8 of experiment - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 10//) __Wednesday, 11th__ - Conduct Day 9 of experiment - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 11//) - Note: Based from previous day's results the later process of experiment may be altered - If tomorrow's culture show red, need to consult alternate protocols __Thursday, 12th__ - Observe the bacteria samples and confirm all the streaked samples had red color - Conclude that all of the colonies did not have the desired combination of DNA plasmid - Discuss possible plans of conducting second method of determining the plasmid combination - Discuss with laser cutting technician of preparation of CAD drawing - Technician also approved of design - Will plan to laser cut tomorrow __Friday, 13th__ - Finalize AutoCAD drawing before laser cutting - Instead of creating spaces between parts, place parts close together to save plastic - Cut 19/23 parts required for our SDS page model - Please check below link to view photographs of the laser cutting - Go to [[start:classes:principlesofdesign:igem:photographs|Lab Photos]] - Create initial assembly by using tape - Will cut the remaining pieces next Monday and plan on gluing __Monday, 16th__ - Laser cut remaining four pieces of SDS page model - Create mock assembly with all of the parts using tape - Glue all of 23 parts for SDS page at the Materials Science Lab - Please consult the below link to view the photographs of the model - Go to [[start:classes:principlesofdesign:igem:photographs|Lab Photos]] - Will test for leak points in the upcoming days __Friday, 20th__ - Pour water in the two chambers of the SDS page to check for leaking points - Determine two potential leaking points on top chamber - Tape the leaking points as markers - Will let the SDS page dry over the weekend before regluing the leaking spots __Monday, 23rd__ - Conduct Day 10 of experiment, restriction digest (following method 2) - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 12//) - Glue the weak spots on the SDS page - Measure and cut spacers that will be used - Place plugs into respective holes - Place wire in the top and bottom chambers - Will test and run gel electrophoresis in the upcoming day or two __Tuesday, 24th__ - Begin gluing second model of the SDS page - Conduct Day 11 of experiment, DNA ligation - Please check the lab work section for the exact procedure - Go to [[start:classes:principlesofdesign:igem:lab_work|Lab Work]] (//Note: Start at Day 13//) - Create 1% agarose gel - Reproduce testing conditions for SDS page - Instead of two different buffers, use one singe TAE buffer for top and bottom chambers - Attempt testing-could not conduct test because gel slipped with the addition of buffer - Need to redefine model and research existing models to solve major problem - Leaking problem is completely solved __Thursday, 26th__ - Research existing models and look at patents - Redefine the existence of porous gel underneath the testing agarose gel - For next SolidWorks model will include the following modifications: * Include plastic shelving to the sides of chamber for a more stable wiring, thus uniform current * Include a mini-chamber underneath to place an agarose gel to prevent slipping of gel __Monday, 30th__ - Laser cut remaining, refinement pieces to SDS page model (bottom stopper-1/4 inch) - Create 1% agarose gel - Prototype testing - no major leaks (after inserting TAE buffer to both chambers) - Gel did not slip - Pipette test dye - Current goes through - separation apparent (successful test) - Please check the lab photographs section for detailed pictures of the prototype test - Go to [[start:classes:principlesofdesign:igem:photographs|Lab Photos]] Back to [[start:classes:principlesofdesign:igem:start|iGEM]]