Experiment: |
Name(s) |
Date |
| Miniprep of yeast backbones from E. coli culture (pRS313, pRS315, pRS316) |
Everyone (and the high schoolers) |
6/15/2015 |
| Preparation of competent E. coli |
Mark, Lexi |
6/19/2015 |
| Transformation of backbones into yeast culture |
Tony, Bo |
6/22/2015 |
| 3A Assembly of killswitch promoter to RBS |
Everyone |
6/23/2015 |
| Transformation of GFP into E. coli to test competency of cells we made ourselves |
Bo, Lexi |
6/24/2015 |
| Digestion of promoter, rbs, lysing agent, and terminator |
Bo, Lexi, Kate |
6/24/2015 |
| Ligation and transformation of promoter rbs lysing agent and terminator |
Bo, Lexi, Kate |
6/25/2015 |
| PCR of G-Block Fragments |
Mark, Melissa, Erich, Suraj, Jill |
6/25/2015 |
| |
|
//2015 |
| Resuspension and Dilution of G-Blocks and Primers |
Mark, Melissa, Erich, Suraj, Jill |
6/25/2015 |
| PCR (3 replicates) of LiP1 and LiP2 |
Mark, Melissa, Erich, Suraj, Jill |
6/26/2015 |
| PCR (3 replicates) of Lac1 and Lac2 |
Mark, Melissa, Erich, Jill |
6/26/2015 |
| Run a gel to check digestion of promoter rbs lysing agent and terminator |
Bo, Lexi, Kate |
6/26/2015 |
| PCR (3 replicates) of MnP1 and MnP2 |
Mark, Melissa, Jill |
6/29/2015 |
| PCR (3 replicates) of AKR1, VP1, VP2 |
Melissa |
6/30/2015 |
| PCR (3 replicates) of AKR2, Tyr1, Tyr2 |
Melissa, Erich |
7/1/2015 |
| Gel of Lac1, Lac2, MnP1, MnP2, VP1 |
Mark, Jill |
7/1/2015 |
| PCR (3 replicates) of all gBlocks |
Mark, Melissa, Jill, Erich |
7/1/2015 |
| Miniprep of promoter, rbs, lysing agent, and terminator |
Bo, Lexi, Kate |
7/6/2015 |
| Digestion of promoter with rbs and lysing agent, with terminator |
Bo, Lexi, Kate |
7/7/2015 |
| PCR Purification (of MnP1&2, AKR1&2, VP1&2, and Tyr1) |
Suraj, Melissa |
7/7/2015 |
| PCR of all G-Block Fragments |
Jill, Mark, Erich |
7/7/2015 |
| Inoculation of liquid yeast cultures |
Mark |
7/7/2015 |
| LB Agar with Amp plates |
Bo, Lexi, Kate |
7/8/2015 |
| Gel of LiP1, LiP2, Lac1, Lac2, Tyr2 |
Mark, Melissa, Erich, Suraj, Jill |
7/8/2015 |
| Gibson Assembly with PCR purified gBlocks (Mn1+Mn2, AKR1+AKR2, VP1+VP2) |
Mark |
7/8/2015 |
| Gel of LiP1, LiP2, Lac1, Lac2, Tyr2 (of second PCR attempt) |
Mark, Melissa, Erich, Suraj, Jill |
7/8/2015 |
| PCR Purification of LiP1 and LiP2 |
Jill, Erich, Melissa, Suraj |
7/8/2015 |
| Gibson Assembly with PCR purified gBlocks (LiP1 + LiP2) |
Jill, Erich, Melissa, Suraj |
7/8/2015 |
| E. Coli Transformation of Promoter-RBS and LysingAgent-Terminator Ligations |
Bo, Lexi, Kate |
7/8/2015 |
| Yeast Transformation of backbones pRS313, pRS315, and pRS316 |
Mark, Tony |
7/8/2015 |
| Gel of Gibson Results LiP, MnP, AKR, VP |
Erich, Suraj, Melissa |
7/9/2015 |
| LB broth with amp |
Bo, Lexi, Kate |
7/9/2015 |
| Ligation of promoter+rbs and lysing agent+terminator |
Bo, Lexi, Kate |
7/9/2015 |
| E. Coli Transformation of Promoter-RBS and LysingAgent-Terminator Ligations |
Bo, Lexi, Kate |
7/10/2015 |
| Gibson Assembly of PCR purified VP1 + VP2 |
Jill, Erich, Melissa, Suraj |
7/10/2015 |
| Gel of Gibson Results VP |
Jill, Erich, Melissa, Suraj |
7/10/2015 |
| Ethanol Precipitation of VP1 and VP2 |
Jill, Erich, Melissa, Suraj, Mark |
7/10/2015 |
| Non-inoculated yeast plates test |
Tony |
7/10/2015 |
| Inoculation of Promoter+RBS and LysingAgent+Terminator E. Coli |
Bo, Lexi, Kate |
7/12/2015 |
| Preparation of Yeast glycerol stocks for long-term storage |
Mark, Erich |
7/13/2015 |
| Miniprep of promoter+rbs and lysing agent+terminator |
Bo, Lexi, Kate |
7/13/2015 |
| Gibson Assembly of Ethanol Precipitated VP1+VP2 |
Jill, Erich, Melissa, Suraj |
7/13/2015 |
| Gel of Gibson Results VP |
Jill, Erich, Melissa, Suraj |
7/13/2015 |
| Inoculation of liquid yeast cultures |
Mark |
7/13/2015 |
| Second inoculation of liquid yeast cultures for competent cell preparation |
Mark |
7/14/2015 |
| Transformation of second diluted yeast cultures with pRS313 |
Mark, Jill, Erich, Melissa, Suraj |
7/14/2015 |
| Inoculation of liquid yeast cultures |
Mark |
7/14/2015 |
| Dilution (10x, 25x, 50x, 100x) of inoculated yeast cultures |
Mark |
7/15/2015 |
| Miniprep of promoter+rbs and lysing agent+terminator |
Bo, Lexi, Kate |
7/15/2015 |
| Digestion of promoter+rbs and lysing agent+terminator |
Bo, Lexi, Kate |
7/15/2015 |
| Preparation of E. Coli glycerol stocks for P+R and L+T |
Bo, Lexi, Kate |
7/15/2015 |
| Overlap PCR of MnP1 and 2 (w/ primers and w/o) |
Mark, Jill, Erich, Melissa, Suraj |
7/15/2015 |
| Gel of PCR Overlap Results |
Jill, Erich, Melissa, Suraj |
7/15/2015 |
| Ligation of promoter+rbs+lysing agent+terminator |
Bo, Lexi, Kate |
7/15/2015 |
| Transformation of promoter+rbs+lysing agent+terminator |
Bo, Kate, Lexi |
7/16/2015 |
| Gel Electrophoresis of MnP Overlap PCR |
Mark, Suraj |
7/16/2015 |
| Overlap PCR of LiP (straight gBlocks and PCR amplified gBlocks) |
Mark, Suraj |
7/16/2015 |
| Gel Extraction of MnP |
Erich |
7/16/2015 |
| Overlap PCR of LiP (straight gBlocks) |
Suraj, Tony, Arren, Jill, James |
7/16/2015 |
| Gel of Overlap PCR of LiP |
Suraj, Jill, Melissa, Erich |
7/17/2015 |
| PCR Amplification of Lac1, Lac2, Tyr2 |
Suraj, Jill, Melissa |
7/20/2015 |
| Overlap PCR Part 2 of MnP |
Erich |
7/20/2015 |
| Overlap PCR (with calculated Tm) of VP |
Suraj, Jill, Melissa |
7/20/2015 |
| Ligation of promoter+rbs+lysing agent+terminator |
Bo, Lexi, Kate |
7/20/2015 |
| Overlap PCR (with calculated Tm) of AKR |
Suraj, Melissa, Erich |
7/21/2015 |
| Restriction Enzyme Digest and Ligation of MnP to pRS315 |
Mark, Erich |
7/21/2015 |
| LB w/ chlor made |
Bo, Kate, Lexi |
7/21/2015 |
| Inoculation of full device and E. coli |
Bo, Kate, Lexi |
7/21/2015 |
PCR Amplification of LiP1, LiP2, Lac1, Lac2, VP1, VP2, AKR1, AKR2, Tyr1, Tyr2 |
Melissa, Jill |
7/21/2015 |
| E. coli transformation of MnP + pRS315 |
Mark |
7/22/2015 |
| Gel of PCR Products (LiP1, LiP2, Lac1, Lac2, VP1, VP2, AKR1, AKR2, Tyr1, Tyr2) |
Suraj, Erich |
7/22/2015 |
| Gibson assembly of AKR1 + AKR 2 |
Jill |
7/22/2015 |
| Gel of Gibson Results AKR |
Suraj |
7/22/2015 |
| PCR Purification of AKR2, LiP2, VP1, VP2 |
Jill |
7/22/2015 |
| Mini prep + nanodrop |
Bo, Lexi, Kate |
7/23/2015 |
| Gibson Assembly of VP1 + VP2 |
Mark |
7/23/2015 |
| Gel Extraction of LiP1, Lac1, Lac2, AKR1, |
Suraj, Melissa, Jill |
7/23/2015 |
| Gel Tyr1, Tyr2, Lac2, Gibson Assembley of AKR |
Suraj, Tony |
7/23/2015 |
| Streaking of new Yeast (BY4741) plate |
Mark |
7/23/2015 |
| EcoRI and PstI digest of AKR and MnP |
Mark |
7/27/2015 |
Liquid Inoculation of Yeast MST WT Cultures |
Erich |
7/27/2015 |
| Digestion of GFP and full device for insertion into chlor and amp backbone |
Lexi, Kate |
7/27/2015 |
| PCR of Lac2 and Gibson AKR |
Tony, Melissa |
7/27/2015 |
| Gel electrophoresis of Lac2 (for extraction), AKR(test), & Tyr1(for extraction) |
Tony, Melissa |
7/27/2015 |
| Vector Insertion of AKR and MnP into pSCB13C |
Mark |
7/27/2015 |
| Transformation of AKR and MnP and GFP |
Bo, Kate, Lexi |
7/28/2015 |
| Yeast Transformation of backbones pRS313, pRS315, and pRS316 |
Erich |
7/28/2015 |
| LB + Chlor plates made |
Tony |
7/28/2015 |
| Gel of Lac1 and Lac2 PCR amplified products (140uL in big wells) |
Suraj, Melissa |
7/29/2015 |
| PCR Purification of LiP Gibson Results |
Jill, Melissa |
7/29/2015 |
| Digestion of Full Device and yeast backbone |
Bo, Lexi, Kate |
8/3/2015 |
| Digest of Mnp, AKR, GFP, full device and backbone |
Bo, Kate, Lexi |
8/3/2015 |
EtBr Gel of (Lane1: Ladder, Lane2: Tyr Gibson 20uL, Lane3: PCRLac1 A 10uL, Lane4: Lac1B, Lane5: Lac1C, Lane6: Lac2A, Lane7: Lac2B, Lane8: Lac2C) |
Jill, Erich, Suraj, Melissa |
8/3/2015 |
| Transformation of AKR, MnP, GFP, and full device |
Bo, Kate, Lexi |
8/4/2015 |
| Gel of Lac1 and Lac2 PCR amplified products (140uL in big wells) (Lane 1: Ladder, Lane 2: Lac1, Lane3: Lac2) |
Jill, Erich, Suraj, Melissa |
8/4/2015 |
| Gel of (Lane1: Ladder, Lane2: 5ul Tyr 1 Gel extract, Lane3: 5uL Tyr2 Gel Extract) |
Jill, Erich, Suraj, Melissa |
8/4/2015 |
| Gel Extraction of PCR amplified Lac1 and Lac2 |
Jill, Suraj, Erich |
8/4/2015 |
| Gibson Assembly of Tyr1+Tyr2 (different gel extraction than before) |
Jill, Melissa |
8/4/2015 |
| Gibson Assembly of Lac1 + Lac2 gel extracts |
Jill, Melissa |
8/5/2015 |
| Gel of Gibson Assembly results (Lane1: Ladder, Lane2: Tyr, Lane3: Lac) |
Jill, Suraj, Melissa |
8/5/2015 |
| Gibson Assembly of Tyr1+2 and Lac1+2 (with more DNA added) |
Jill, Suraj, Melissa |
8/5/2015 |
| Gel Extract of Gibson Assembly Results Tyr |
Erich |
8/5/2015 |
| Re-Gibson Assembly of Lac1 & Lac2 |
Jill, Suraj |
8/6/2015 |
| PCR of Tyr |
Jill, Suraj |
8/6/2015 |
Gel of (Lane1: Ladder, Lane 2: Lac Gibson, Lane 3: Lac Gibson, Lane 4: Tyr1, Lane5: Tyr2, Lane6: Tyr3, empty, empty) |
Jill, Suraj |
8/6/2015 |
| Gel of the rest of Tyr (140uL) (Lane 1: ladder, Lane2: Tyr, empty) |
Jill, Suraj |
8/6/2015 |
| Gel Extraction of Lac Gibson (3.5ng/uL) |
Jill, Suraj |
8/7/2015 |
| Re-measurement of Lac and Tyr concentrations using Cubit instead of nanometer |
Jill, Suraj, Melissa |
8/26/2015 |
| PCR of Gibson Tyr and Lac |
Jill, Suraj |
8/27/2015 |
| PCR of Tyr1, Tyr2, Lac1, Lac2 |
Jill, Suraj |
8/27/2015 |
| Gel 1: Lane 1: Ladder, Lane 2: Lac1 PCR, Lane 3: Lac2 PCR, Lane 4 Tyr1 PCR, Lane 5 Tyr2 PCR -----Empty |
Melissa, Jill |
8/28/2015 |
| Gel 2: Lane 1: Ladder, Lane 2: Lac Gibson A, Lane 3: Lac Gibson B, Lane 4 Tyr Gibson A, Lane 5 Tyr Gibson B ---- Empty |
Melissa, Jill |
8/28/2015 |
