Team:Aberdeen Scotland/notebook/andgate
From 2009.igem.org
(Difference between revisions)
(→AND Gate Notebook) |
|||
(14 intermediate revisions not shown) | |||
Line 3: | Line 3: | ||
- | + | =AND Gate Notebook= | |
- | + | ==Week 1: Research and Planning (08/06/09 - 12/06/09)== | |
+ | <br><br> | ||
+ | ===Day 1 Monday (08/06/09)=== | ||
+ | *Researching the Registry for Biobricks | ||
+ | **Focus on T7 parts (I712074, I719005, K113011, K113012, J34814, K103021, R0085, R0180, R0182, Z0251 and Z0252) for possible use in our project. | ||
+ | **Other BioBricks identified for other sub-projects include R0062, J40001, J37015 and R0063 | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (09/06/09)=== | ||
+ | *Researching literature for T7 promoter strength and degradation rate | ||
+ | **Useful article - Imburgio, D., Rong, M., Ma, K., and W. T. McAllister. (2000) "Studies of Promoter Recognition and Start Site Selection by T7 RNA Polymerase Using a Comprehensive Collection of Promoter Variants" Biochemistry.39:10419-10430 | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (10/06/09)=== | ||
- | + | *Researching literature for parameters of LuxR, LacO, TetO and cIO | |
+ | ** Useful article - Braff, J. C., Conboy, C. C., and D.E. Endy. Promoter Characterization Experiments. (found at openwetware.org/images/3/30/PromoterChar_Report_Revised.doc) | ||
+ | *Identified plasmids that potential Biobricks were on and prepared for rescue. | ||
+ | **Also indentified LVA tags and any other anomalies on selected Biobricks. | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (11/06/09)=== | ||
+ | *PoPs to LacZ alpha repoter identified (E0435) and potenially useful Input Output sensor (pSB1A10) | ||
+ | **Other possible reporters include LacZ alpha fragments and Enhanced stable YFP. | ||
+ | *Possibility of intentional SNP generated within T7 to alter promoter strength was deemed unworkable in the scale of project. | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (12/06/09)=== | ||
- | + | *End of week meeting | |
+ | *Discussion and planning for following week | ||
+ | <br><br> | ||
+ | ==Week 2: BioBrick Rescue (15/06/09 - 19/06/09)== | ||
+ | <br><br> | ||
+ | ===Day 1 Monday (15/06/09)=== | ||
- | + | *Prepared LB medium and LB+Amp plates | |
+ | *Preperation of Kanamycin stock (30mg ml-1) | ||
+ | *Inoculated C0040, R0062, J37033 and C0051 for miniprep the following day | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (16/06/09)=== | ||
+ | *Miniprep of above Biobricks | ||
+ | *Restriction digest of C0040, R0062, J37033 and C0051 with EcoRI-HF + SpeI and gel electrophoreis of these. | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (17/06/09)=== | ||
+ | * Rescue of a range of vectors (pSB1AT3, pSB1AC3, pSB3T5, pSB3C5, pSB4T5, pSB4C5) | ||
+ | * Preperation of Tet and Chlo agar plates for the above | ||
+ | * Transformation of DB3.1 cells (ccdB gene resistant) | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (18/06/09)=== | ||
- | === | + | *Preparation of more Chlo and Tet plates. |
+ | *Sub-cultivation of transformants | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (19/06/09)=== | ||
+ | *End of week meeting to present weeks work and plan ahead | ||
+ | <br><br> | ||
+ | ==Week 3: Edinburgh iGEM meeting and Digestions(22/06/09 - 26/06/09)== | ||
+ | ===Day 1 Monday (22/06/09)=== | ||
- | ===Week | + | *Preperation for cloning and selection of desired biobricks from earlier rescues |
+ | <br><br> | ||
+ | ===Day 2 Tuesday (23/06/09)=== | ||
+ | *Edinburgh Igem Meeting | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (24/06/09)=== | ||
+ | *Edinburgh Igem Meeting | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (25/06/09)=== | ||
+ | * Double digest of the biobricks B0030, C0051, I0462, J23105, J23107, J23115, with EcoRI-HF + Spe I | ||
+ | * Double digest of the biobricks S03518, E0840 with XbaI + PstI | ||
+ | * Double digest of the biobricks pSB3K3, pSB4K5, pSB3T5, pSB1AC3, pSB1AT3 | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (26/06/09)=== | ||
+ | *DB3.1 cells transformed with pSB1AK3 | ||
+ | *Gel Electrophoresis of above digests | ||
+ | *End of week meeting | ||
+ | <br><br> | ||
+ | ==Week 4:Digestions Galore(29/06/09 - 03/07/09)== | ||
+ | ===Day 1 Monday (29/06/09)=== | ||
+ | *Dilution double digest of the biobrick E0840 with XbaI + PstI | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (30/06/09)=== | ||
+ | *Double digest of the biobrick C0051 with EcoRI-HF + SpeI | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (01/07/09)=== | ||
+ | *Dilution double digest of pSB3T5 with EcoRI-HF + PstI | ||
+ | *Dilution double digest of E0840 with XbaI + SpeI | ||
+ | *Transformation of DB3.1 competent cells with pSB3K5 | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (02/07/09)=== | ||
+ | *Electrophoresis of the digests form the previous day on 0.8% (E0840) and 1% (C0051) agar | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (03/07/09)=== | ||
+ | *Repeated dilution double digests for biobricks E0840, C0051 and pSB3T5 | ||
+ | *End of week meeting | ||
+ | <br><br> | ||
+ | ==Week 5 Preperation of K182100 (06/07/09 - 11/07/09)== | ||
+ | ===Day 1 Monday (06/07/09)=== | ||
+ | *Planning and calculations for ligation | ||
+ | *Electrophoresis of the dilution double digests | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (07/07/09)=== | ||
+ | *Double digest of the biobrick pSB3K5 | ||
+ | *Dephosphorylation of the pSB3T5 | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (08/07/09)=== | ||
+ | *Electrophoresis of the pSB3K5 double digest | ||
+ | *Electrophoresis of the double digests of J series promoters (J23107, J23105, J23115) | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (09/07/09)=== | ||
+ | *Electrophoresis of the double digest – pSB1AC3 | ||
+ | **Ligation , Cloning and Transformation | ||
+ | **Transformation of XL1 Blue competent cells | ||
+ | *Preparation of IPTG stock | ||
+ | *Preparation of Chlo agar plates | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (10/07/09)=== | ||
+ | *The results of the transformation | ||
+ | *New transformation of the XL1- Blue competent cells with C0051, E0840 and pSB1AC3 | ||
+ | *End of week meeting | ||
+ | <br><br> | ||
- | + | ==Week 6 Identifying K182100 (13/07/09 - 17/07/09)== | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
+ | ===Day 1 Monday (13/07/09)=== | ||
+ | * The results of transformation of XL-1 Blue competent cells with pSB1AC3, C0051, E0840, look positive. | ||
+ | * Preparation of grid plates (with 40 colonies picked from overnight culturem and 5 control colonies from Vector and Vector+Ligase cultures). | ||
+ | *Preparation for PCR colony | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (14/07/09)=== | ||
+ | * Preparation, calculation for PCR colony | ||
+ | * PCR colony | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (15/07/09)=== | ||
+ | * Calculation for double digest of PCR product | ||
+ | * Double digest of PCR product with HindIII and NdeI to hopefully show insert containing part of E0840 and C0051 | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (16/07/09)=== | ||
+ | * Mini prep of good colonies | ||
+ | * Double digest of the positive transformants with EcoRi-HF + PstI and HindIII + NdeI | ||
+ | <br><br> | ||
+ | ===Day 1 Friday (17/07/09)=== | ||
+ | * Weekly meeting, cake and planning ahead | ||
+ | <br><br> | ||
+ | ==Week 7 2nd Cloning Step (20/07/09 - 24/07/09)== | ||
+ | ===Day 1 Monday (20/07/09)=== | ||
+ | * Fusion PCR for K182102 was prepared | ||
+ | * K182100 sent for sequencing | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (21/07/09)=== | ||
+ | * Fusion PCR for K182102 was repeated | ||
+ | * Gel electrophoresis of PCR product | ||
+ | * Gel DNA extraction and purification | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (22/07/09)=== | ||
+ | * Double digest of PCR product and K182100 with XbaI + PstI | ||
+ | * Gel electrophoresis of previous double digests | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (23/07/09)=== | ||
+ | * Calculation for ligation | ||
+ | * Ligation and transformation of SCS1 supercompetent cells | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (24/07/09)=== | ||
+ | * The results form the ligation were obtained, though they showed a poor Vector+Inserts+Ligase ratio to Vector+Ligase. | ||
+ | ** Preparation of a new fusion PCR | ||
+ | ** Gel electrophoresis of PCR product | ||
+ | ** DNA gel extraction and purification | ||
+ | <br><br> | ||
+ | ==Week 8 Déjà vu (27/07/09 - 31/07/09)== | ||
+ | ===Day 1 Monday (27/07/09)=== | ||
+ | * Double digest of PCR product with EcoRI-HF + PstI | ||
+ | * Double digest of PCR product with EcoRI + SpeI | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (28/07/09)=== | ||
+ | * Heat inactivation of restriction enzymes | ||
+ | ** Dephosphorylation of pSB3T5 vector | ||
+ | ** Ligation of pSB3T5 + K182103 | ||
+ | * Motility assay | ||
+ | * Chemotaxis experiment on minimal media with 1%, 2% and 4% aspartate | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (29/07/09)=== | ||
+ | * The results of chemotaxis experiment (unpromising) | ||
+ | * The results of ligation (unpromising) | ||
+ | * Preparation of IPTG | ||
+ | * Preparation of grid plates | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (30/07/09)=== | ||
+ | * Colony PCR | ||
+ | * Double digest of pSB3T5 with EcoRI-HF and PstI | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (31/07/09)=== | ||
+ | * Gel electrophoresis of the pSB3T5 double digest | ||
+ | * End of week meeting | ||
+ | <br><br> | ||
+ | ==Week 9 Motility focus (03/08/09 - 07/08/09)== | ||
+ | ===Day 1 Monday (03/08/09)=== | ||
+ | *New fusion PCR | ||
+ | **Purification of PCR product | ||
+ | ** Double digest and gel electrophoresis | ||
+ | * Motility assay | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (04/08/09)=== | ||
+ | * Calculation for ligation | ||
+ | * Ligation and transformation of SCS-1 supercompetent cells | ||
+ | * Motiliy experiment | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (05/08/09)=== | ||
+ | * The results of ligation and transformation | ||
+ | * Preparation of mini prep | ||
+ | * Motility assay | ||
+ | * Preparation of grid plates | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (06/08/09)=== | ||
+ | * The results of motility assay were obtained | ||
+ | * PCR colony of K182103 | ||
+ | * Gel electrophoresis of the PCR product | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (07/08/09)=== | ||
+ | * Colony PCR of the K182102 | ||
+ | * Gel electrophoresis of the PCR product | ||
+ | * Double digest of the mini preps prepared | ||
+ | <br><br> | ||
+ | ==Week 10 Wrapping Up (10/08/09 - 14/08/09)== | ||
+ | ===Day 1 Monday (10/08/09)=== | ||
+ | * Double digest of the K182100 and potential K182103 | ||
+ | * Estimation of the concentration (ng/ul) in our mini preps to be send for sequencing | ||
+ | * Gel electrophoresis of the double digest | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (11/08/09)=== | ||
+ | * Preparation for the sequencing | ||
+ | * Potential K182102 and K182103 sent for sequencing | ||
+ | * Working on the team wiki | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (12/08/09)=== | ||
+ | * Overnight incubation of the motile strain MG1655 | ||
+ | * Working on the team wiki | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (13/08/09)=== | ||
+ | * Motility and chemotaxis assay, though it failed to yield suitable/usable results | ||
+ | * Results from sequencing K182102, sequencing was satisfactory | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (14/08/09)=== | ||
+ | * Capillary chemotaxis assay | ||
+ | * Weekly meeting and team photos taken | ||
+ | <br><br> | ||
+ | ==Week 11 The End is Nigh! (17/08/09 - 21/08/09)== | ||
+ | ===Day 1 Monday (17/08/09)=== | ||
+ | *Updated wiki notebook | ||
+ | **Added several translations of front page summary to wiki | ||
+ | <br><br> | ||
+ | ===Day 2 Tuesday (18/08/09)=== | ||
+ | *Zuzana - Motility Assay for modelling photographs | ||
+ | *Calum - Had a 30cm plate removed from my right arm and several areas of my shoulderbone cut off | ||
+ | <br><br> | ||
+ | ===Day 3 Wednesday (19/08/09)=== | ||
+ | *Zuzana - Updated wiki Wetlab section | ||
+ | *Calum - Bled on things | ||
+ | <br><br> | ||
+ | ===Day 4 Thursday (20/08/09)=== | ||
+ | *Editing and tidying of wiki and devouring of cake | ||
+ | <br><br> | ||
+ | ===Day 5 Friday (21/08/09)=== | ||
Line 61: | Line 289: | ||
<tr> | <tr> | ||
<td> | <td> | ||
- | <a href=" | + | <a href="https://2009.igem.org/Team:Aberdeen_Scotland/modeling/conclusion"><img src="https://static.igem.org/mediawiki/2009/e/ed/Aberdeen_Left_arrow.png"> Back to Modeling Conclusions</a> |
</td> | </td> | ||
<td align="right"> | <td align="right"> | ||
- | <a href="https://2009.igem.org/Team:Aberdeen_Scotland/notebook/lacilatch">LacI-Latch <img src="https://static.igem.org/mediawiki/2009/4/4c/Aberdeen_Right_arrow.png"></a> | + | <a href="https://2009.igem.org/Team:Aberdeen_Scotland/notebook/lacilatch">Continue to Notebook, LacI-Latch Section <img src="https://static.igem.org/mediawiki/2009/4/4c/Aberdeen_Right_arrow.png"></a> |
</td> | </td> | ||
</tr> | </tr> |
Latest revision as of 09:01, 20 August 2009
University of Aberdeen - Pico Plumber
iGEM 2009
AND Gate Notebook
Week 1: Research and Planning (08/06/09 - 12/06/09)
Day 1 Monday (08/06/09)
- Researching the Registry for Biobricks
- Focus on T7 parts (I712074, I719005, K113011, K113012, J34814, K103021, R0085, R0180, R0182, Z0251 and Z0252) for possible use in our project.
- Other BioBricks identified for other sub-projects include R0062, J40001, J37015 and R0063
Day 2 Tuesday (09/06/09)
- Researching literature for T7 promoter strength and degradation rate
- Useful article - Imburgio, D., Rong, M., Ma, K., and W. T. McAllister. (2000) "Studies of Promoter Recognition and Start Site Selection by T7 RNA Polymerase Using a Comprehensive Collection of Promoter Variants" Biochemistry.39:10419-10430
Day 3 Wednesday (10/06/09)
- Researching literature for parameters of LuxR, LacO, TetO and cIO
- Useful article - Braff, J. C., Conboy, C. C., and D.E. Endy. Promoter Characterization Experiments. (found at openwetware.org/images/3/30/PromoterChar_Report_Revised.doc)
- Identified plasmids that potential Biobricks were on and prepared for rescue.
- Also indentified LVA tags and any other anomalies on selected Biobricks.
Day 4 Thursday (11/06/09)
- PoPs to LacZ alpha repoter identified (E0435) and potenially useful Input Output sensor (pSB1A10)
- Other possible reporters include LacZ alpha fragments and Enhanced stable YFP.
- Possibility of intentional SNP generated within T7 to alter promoter strength was deemed unworkable in the scale of project.
Day 5 Friday (12/06/09)
- End of week meeting
- Discussion and planning for following week
Week 2: BioBrick Rescue (15/06/09 - 19/06/09)
Day 1 Monday (15/06/09)
- Prepared LB medium and LB+Amp plates
- Preperation of Kanamycin stock (30mg ml-1)
- Inoculated C0040, R0062, J37033 and C0051 for miniprep the following day
Day 2 Tuesday (16/06/09)
- Miniprep of above Biobricks
- Restriction digest of C0040, R0062, J37033 and C0051 with EcoRI-HF + SpeI and gel electrophoreis of these.
Day 3 Wednesday (17/06/09)
- Rescue of a range of vectors (pSB1AT3, pSB1AC3, pSB3T5, pSB3C5, pSB4T5, pSB4C5)
- Preperation of Tet and Chlo agar plates for the above
- Transformation of DB3.1 cells (ccdB gene resistant)
Day 4 Thursday (18/06/09)
- Preparation of more Chlo and Tet plates.
- Sub-cultivation of transformants
Day 5 Friday (19/06/09)
- End of week meeting to present weeks work and plan ahead
Week 3: Edinburgh iGEM meeting and Digestions(22/06/09 - 26/06/09)
Day 1 Monday (22/06/09)
- Preperation for cloning and selection of desired biobricks from earlier rescues
Day 2 Tuesday (23/06/09)
- Edinburgh Igem Meeting
Day 3 Wednesday (24/06/09)
- Edinburgh Igem Meeting
Day 4 Thursday (25/06/09)
- Double digest of the biobricks B0030, C0051, I0462, J23105, J23107, J23115, with EcoRI-HF + Spe I
- Double digest of the biobricks S03518, E0840 with XbaI + PstI
- Double digest of the biobricks pSB3K3, pSB4K5, pSB3T5, pSB1AC3, pSB1AT3
Day 5 Friday (26/06/09)
- DB3.1 cells transformed with pSB1AK3
- Gel Electrophoresis of above digests
- End of week meeting
Week 4:Digestions Galore(29/06/09 - 03/07/09)
Day 1 Monday (29/06/09)
- Dilution double digest of the biobrick E0840 with XbaI + PstI
Day 2 Tuesday (30/06/09)
- Double digest of the biobrick C0051 with EcoRI-HF + SpeI
Day 3 Wednesday (01/07/09)
- Dilution double digest of pSB3T5 with EcoRI-HF + PstI
- Dilution double digest of E0840 with XbaI + SpeI
- Transformation of DB3.1 competent cells with pSB3K5
Day 4 Thursday (02/07/09)
- Electrophoresis of the digests form the previous day on 0.8% (E0840) and 1% (C0051) agar
Day 5 Friday (03/07/09)
- Repeated dilution double digests for biobricks E0840, C0051 and pSB3T5
- End of week meeting
Week 5 Preperation of K182100 (06/07/09 - 11/07/09)
Day 1 Monday (06/07/09)
- Planning and calculations for ligation
- Electrophoresis of the dilution double digests
Day 2 Tuesday (07/07/09)
- Double digest of the biobrick pSB3K5
- Dephosphorylation of the pSB3T5
Day 3 Wednesday (08/07/09)
- Electrophoresis of the pSB3K5 double digest
- Electrophoresis of the double digests of J series promoters (J23107, J23105, J23115)
Day 4 Thursday (09/07/09)
- Electrophoresis of the double digest – pSB1AC3
- Ligation , Cloning and Transformation
- Transformation of XL1 Blue competent cells
- Preparation of IPTG stock
- Preparation of Chlo agar plates
Day 5 Friday (10/07/09)
- The results of the transformation
- New transformation of the XL1- Blue competent cells with C0051, E0840 and pSB1AC3
- End of week meeting
Week 6 Identifying K182100 (13/07/09 - 17/07/09)
Day 1 Monday (13/07/09)
- The results of transformation of XL-1 Blue competent cells with pSB1AC3, C0051, E0840, look positive.
- Preparation of grid plates (with 40 colonies picked from overnight culturem and 5 control colonies from Vector and Vector+Ligase cultures).
- Preparation for PCR colony
Day 2 Tuesday (14/07/09)
- Preparation, calculation for PCR colony
- PCR colony
Day 3 Wednesday (15/07/09)
- Calculation for double digest of PCR product
- Double digest of PCR product with HindIII and NdeI to hopefully show insert containing part of E0840 and C0051
Day 4 Thursday (16/07/09)
- Mini prep of good colonies
- Double digest of the positive transformants with EcoRi-HF + PstI and HindIII + NdeI
Day 1 Friday (17/07/09)
- Weekly meeting, cake and planning ahead
Week 7 2nd Cloning Step (20/07/09 - 24/07/09)
Day 1 Monday (20/07/09)
- Fusion PCR for K182102 was prepared
- K182100 sent for sequencing
Day 2 Tuesday (21/07/09)
- Fusion PCR for K182102 was repeated
- Gel electrophoresis of PCR product
- Gel DNA extraction and purification
Day 3 Wednesday (22/07/09)
- Double digest of PCR product and K182100 with XbaI + PstI
- Gel electrophoresis of previous double digests
Day 4 Thursday (23/07/09)
- Calculation for ligation
- Ligation and transformation of SCS1 supercompetent cells
Day 5 Friday (24/07/09)
- The results form the ligation were obtained, though they showed a poor Vector+Inserts+Ligase ratio to Vector+Ligase.
- Preparation of a new fusion PCR
- Gel electrophoresis of PCR product
- DNA gel extraction and purification
Week 8 Déjà vu (27/07/09 - 31/07/09)
Day 1 Monday (27/07/09)
- Double digest of PCR product with EcoRI-HF + PstI
- Double digest of PCR product with EcoRI + SpeI
Day 2 Tuesday (28/07/09)
- Heat inactivation of restriction enzymes
- Dephosphorylation of pSB3T5 vector
- Ligation of pSB3T5 + K182103
- Motility assay
- Chemotaxis experiment on minimal media with 1%, 2% and 4% aspartate
Day 3 Wednesday (29/07/09)
- The results of chemotaxis experiment (unpromising)
- The results of ligation (unpromising)
- Preparation of IPTG
- Preparation of grid plates
Day 4 Thursday (30/07/09)
- Colony PCR
- Double digest of pSB3T5 with EcoRI-HF and PstI
Day 5 Friday (31/07/09)
- Gel electrophoresis of the pSB3T5 double digest
- End of week meeting
Week 9 Motility focus (03/08/09 - 07/08/09)
Day 1 Monday (03/08/09)
- New fusion PCR
- Purification of PCR product
- Double digest and gel electrophoresis
- Motility assay
Day 2 Tuesday (04/08/09)
- Calculation for ligation
- Ligation and transformation of SCS-1 supercompetent cells
- Motiliy experiment
Day 3 Wednesday (05/08/09)
- The results of ligation and transformation
- Preparation of mini prep
- Motility assay
- Preparation of grid plates
Day 4 Thursday (06/08/09)
- The results of motility assay were obtained
- PCR colony of K182103
- Gel electrophoresis of the PCR product
Day 5 Friday (07/08/09)
- Colony PCR of the K182102
- Gel electrophoresis of the PCR product
- Double digest of the mini preps prepared
Week 10 Wrapping Up (10/08/09 - 14/08/09)
Day 1 Monday (10/08/09)
- Double digest of the K182100 and potential K182103
- Estimation of the concentration (ng/ul) in our mini preps to be send for sequencing
- Gel electrophoresis of the double digest
Day 2 Tuesday (11/08/09)
- Preparation for the sequencing
- Potential K182102 and K182103 sent for sequencing
- Working on the team wiki
Day 3 Wednesday (12/08/09)
- Overnight incubation of the motile strain MG1655
- Working on the team wiki
Day 4 Thursday (13/08/09)
- Motility and chemotaxis assay, though it failed to yield suitable/usable results
- Results from sequencing K182102, sequencing was satisfactory
Day 5 Friday (14/08/09)
- Capillary chemotaxis assay
- Weekly meeting and team photos taken
Week 11 The End is Nigh! (17/08/09 - 21/08/09)
Day 1 Monday (17/08/09)
- Updated wiki notebook
- Added several translations of front page summary to wiki
Day 2 Tuesday (18/08/09)
- Zuzana - Motility Assay for modelling photographs
- Calum - Had a 30cm plate removed from my right arm and several areas of my shoulderbone cut off
Day 3 Wednesday (19/08/09)
- Zuzana - Updated wiki Wetlab section
- Calum - Bled on things
Day 4 Thursday (20/08/09)
- Editing and tidying of wiki and devouring of cake
Day 5 Friday (21/08/09)
Back to Modeling Conclusions | Continue to Notebook, LacI-Latch Section |