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Team:VictoriaBC/Labnotebook
From 2009.igem.org
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Jasoncummer (Talk | contribs) |
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| Line 62: | Line 62: | ||
New DH5alpha plates | New DH5alpha plates | ||
| - | Lab July 17 2009 | + | [[Lab July 17 2009]] |
| Line 68: | Line 68: | ||
Acheived DH5α growth | Acheived DH5α growth | ||
| - | Lab July 20 2009 | + | [[Lab July 20 2009]] |
Make competent cells following top-10 protocol | Make competent cells following top-10 protocol | ||
| - | Lab July 21 2009 | + | [[Lab July 21 2009]] |
Measurement of competence | Measurement of competence | ||
| - | Lab July 22 2009 | + | [[Lab July 22 2009]] |
Failed competence, started on BCMB301 competent cells protocol | Failed competence, started on BCMB301 competent cells protocol | ||
| - | Lab July 23 2009 | + | [[Lab July 23 2009]] |
Test of competence on new cells, greater quantity of pUC18 | Test of competence on new cells, greater quantity of pUC18 | ||
| - | Lab July 24 2009 | + | [[Lab July 24 2009]] |
| Line 90: | Line 90: | ||
Successful test of competence with pUC18 | Successful test of competence with pUC18 | ||
| - | Lab July 27 2009 | + | [[Lab July 27 2009]] |
Initial messing with rehydration protocol | Initial messing with rehydration protocol | ||
| - | Lab July 28 2009 | + | [[Lab July 28 2009]] |
Successful biobrick transformation | Successful biobrick transformation | ||
| - | Lab July 29 2009 | + | [[Lab July 29 2009]] |
Nandrop rehydrations, more transformations | Nandrop rehydrations, more transformations | ||
| - | Lab July 30 2009 | + | [[Lab July 30 2009]] |
Failed lambda cI promoter | Failed lambda cI promoter | ||
| - | Lab July 31 2009 | + | [[Lab July 31 2009]] |
| Line 112: | Line 112: | ||
Checked weekend culture of RFP | Checked weekend culture of RFP | ||
| - | Lab Aug 4 2009 | + | [[Lab Aug 4 2009]] |
Another nanodrop, more rehydrations and transformations, ran a gel (B0015 Amp/Kan, J06504 Amp, R0051 Amp, B0034 Amp) | Another nanodrop, more rehydrations and transformations, ran a gel (B0015 Amp/Kan, J06504 Amp, R0051 Amp, B0034 Amp) | ||
| - | Lab Aug 5 2009 | + | [[Lab Aug 5 2009]] |
More transformations, struggling with I0500 | More transformations, struggling with I0500 | ||
| - | Lab Aug 6 2009 | + | [[Lab Aug 6 2009]] |
Struggling with Kan growth, more nanodrop rehydrations | Struggling with Kan growth, more nanodrop rehydrations | ||
| - | Lab Aug 7 2009 | + | [[Lab Aug 7 2009]] |
| Line 130: | Line 130: | ||
Tried to induce high copy for I0500 using IPTG | Tried to induce high copy for I0500 using IPTG | ||
| - | Lab Aug 10 2009 | + | [[Lab Aug 10 2009]] |
Make ccdB competent cells, miniprep plasmids, more transformations | Make ccdB competent cells, miniprep plasmids, more transformations | ||
| - | Lab Aug 11 2009 | + | [[Lab Aug 11 2009]] |
Try again at making ccdb strains with Kan and Cm resistances, more transformations | Try again at making ccdb strains with Kan and Cm resistances, more transformations | ||
| - | Lab Aug 12 2009 | + | [[Lab Aug 12 2009]] |
Check on ccdB strains, minipreps, etc. | Check on ccdB strains, minipreps, etc. | ||
| - | Lab Aug 13 2009 | + | [[Lab Aug 13 2009]] |
Make new ccdB competent cells, check on cultures, miniprep, gels (P1010 Kan, J23102, J45120, J45200, K145303, P0151, I0500, C0083, E0422, R1051) | Make new ccdB competent cells, check on cultures, miniprep, gels (P1010 Kan, J23102, J45120, J45200, K145303, P0151, I0500, C0083, E0422, R1051) | ||
| - | Lab Aug 14 2009 | + | [[Lab Aug 14 2009]] |
| Line 152: | Line 152: | ||
Brainstorming, trying to transform P1010 | Brainstorming, trying to transform P1010 | ||
| - | Lab Aug 17 2009 | + | [[Lab Aug 17 2009]] |
Possible success in transforming P1010 | Possible success in transforming P1010 | ||
| - | Lab Aug 18 2009 | + | [[Lab Aug 18 2009]] |
Miniprep and check plasmids, cut and ligated and transformed first assemblies, a few broth cultures, gel (P1010 Kan, P1010 Cm C) | Miniprep and check plasmids, cut and ligated and transformed first assemblies, a few broth cultures, gel (P1010 Kan, P1010 Cm C) | ||
| - | Lab Aug 19 2009 | + | [[Lab Aug 19 2009]] |
Transformed second assemblies, gel (digests) | Transformed second assemblies, gel (digests) | ||
| - | Lab Aug 20 2009 | + | [[Lab Aug 20 2009]] |
Gels (K235000, K235001 and K235002) | Gels (K235000, K235001 and K235002) | ||
| - | Lab Aug 21 2009 | + | [[Lab Aug 21 2009]] |
| Line 174: | Line 174: | ||
Gel (linearized DNA) | Gel (linearized DNA) | ||
| - | Lab Aug 24 2009 | + | [[Lab Aug 24 2009]] |
Gels and assemblies (K145303 better) | Gels and assemblies (K145303 better) | ||
| - | Lab Aug 25 2009 | + | [[Lab Aug 25 2009]] |
Pour Cm plates | Pour Cm plates | ||
| - | Lab Aug 26 2009 | + | [[Lab Aug 26 2009]] |
Re-plate ligation products, new competent cells, failed 2% gels (digests) | Re-plate ligation products, new competent cells, failed 2% gels (digests) | ||
| - | Lab Aug 27 2009 | + | [[Lab Aug 27 2009]] |
Re-plate remaining assemblies, start fridge and freezer cleanup, gels (digests) | Re-plate remaining assemblies, start fridge and freezer cleanup, gels (digests) | ||
| - | Lab Aug 28 2009 | + | [[Lab Aug 28 2009]] |
Brainstorming | Brainstorming | ||
| - | Lab Aug 29, 2009 | + | [[Lab Aug 29, 2009]] |
| Line 202: | Line 202: | ||
Plan for the week, gels (?) | Plan for the week, gels (?) | ||
| - | Lab Aug 31 2009 | + | [[Lab Aug 31 2009]] |
Check P1010Kan, broth culture, gel (K081005, K098988) | Check P1010Kan, broth culture, gel (K081005, K098988) | ||
| - | Lab Sept 01 2009 | + | [[Lab Sept 01 2009]] |
Reassembling K235003, 05 and 10, starting J04450, plating stabs, talking to many people | Reassembling K235003, 05 and 10, starting J04450, plating stabs, talking to many people | ||
| - | Lab Sept 02 2009 | + | [[Lab Sept 02 2009]] |
Broth cultures | Broth cultures | ||
| - | Lab Sept 3 2009 | + | [[Lab Sept 3 2009]] |
Lab move, miniprep and gels (K235003 Cm, K235005 Cm, K235010 Cm, J04450 Kan (+IPTG), J04450 Kan (-IPTG), P1010 Kan, P1010 Amp, R0051 Amp, I0500 Kan (+IPTG), I0500 Kan (-IPTG)) | Lab move, miniprep and gels (K235003 Cm, K235005 Cm, K235010 Cm, J04450 Kan (+IPTG), J04450 Kan (-IPTG), P1010 Kan, P1010 Amp, R0051 Amp, I0500 Kan (+IPTG), I0500 Kan (-IPTG)) | ||
| - | Lab Sep 4 2009 | + | [[Lab Sep 4 2009]] |
| Line 224: | Line 224: | ||
Scheduling a lab meetup for Wednesday, some assemblies, a subculture | Scheduling a lab meetup for Wednesday, some assemblies, a subculture | ||
| - | Lab Sept 08 2009 | + | [[Lab Sept 08 2009]] |
Talk-talk and resources | Talk-talk and resources | ||
| - | Lab Sep 9 2009 | + | [[Lab Sep 9 2009]] |
Transformations | Transformations | ||
| - | Lab Sep 10 2009 | + | [[Lab Sep 10 2009]] |
Broth cultures and rehydrations | Broth cultures and rehydrations | ||
| - | Lab Sep 11 2009 | + | [[Lab Sep 11 2009]] |
Minipreps and broth cultures | Minipreps and broth cultures | ||
| - | Lab Sep 12 2009 | + | [[Lab Sep 12 2009]] |
| Line 246: | Line 246: | ||
Aborted miniprep | Aborted miniprep | ||
| - | Lab Sep 14 2009 | + | [[Lab Sep 14 2009]] |
Broth cultures | Broth cultures | ||
| - | Lab Sep 15 2009 | + | [[Lab Sep 15 2009]] |
Minipreps | Minipreps | ||
| - | Lab Sep 16 2009 | + | [[Lab Sep 16 2009]] |
Gels (K235009, C0012, K235013, K115017, K235014, E0240, K235016, K235017, R0051) | Gels (K235009, C0012, K235013, K115017, K235014, E0240, K235016, K235017, R0051) | ||
| - | Lab Sep 17 2009 | + | [[Lab Sep 17 2009]] |
Flourescence microscopy - failed | Flourescence microscopy - failed | ||
| - | Lab Sep 18 2009 | + | [[Lab Sep 18 2009]] |
| Line 268: | Line 268: | ||
Assemblies, transformations | Assemblies, transformations | ||
| - | Lab Sep 22 2009 | + | [[Lab Sep 22 2009]] |
Broth cultures not done | Broth cultures not done | ||
| - | Lab Sep 23 2009 | + | [[Lab Sep 23 2009]] |
Broth cultures | Broth cultures | ||
| - | Lab Sep 24 2009 | + | [[Lab Sep 24 2009]] |
Miniprep, gels (failed) | Miniprep, gels (failed) | ||
| - | Lab Sep 25 2009 | + | [[Lab Sep 25 2009]] |
Subcultures | Subcultures | ||
| - | Lab Sep 27 2009 | + | [[Lab Sep 27 2009]] |
| Line 290: | Line 290: | ||
Check cultures | Check cultures | ||
| - | Lab Sep 28 2009 | + | [[Lab Sep 28 2009]] |
| Line 296: | Line 296: | ||
Transformations | Transformations | ||
| - | Lab Oct 8 2009 | + | [[Lab Oct 8 2009]] |
Broth cultures and transformations | Broth cultures and transformations | ||
| - | Lab Oct 9 2009 | + | [[Lab Oct 9 2009]] |
Minipreps and gel (K235019 Amp, K235020 Amp, K235021 Amp, K115017 Amp, C0012 Amp, J04630 Amp, J23070 Amp, R0051 Amp) | Minipreps and gel (K235019 Amp, K235020 Amp, K235021 Amp, K115017 Amp, C0012 Amp, J04630 Amp, J23070 Amp, R0051 Amp) | ||
| - | Lab Oct 10 2009 | + | [[Lab Oct 10 2009]] |
Minipreps | Minipreps | ||
| - | Lab Oct 11 2009 | + | [[Lab Oct 11 2009]] |
| Line 314: | Line 314: | ||
Broth cultures | Broth cultures | ||
| - | Lab Oct 12 2009 | + | [[Lab Oct 12 2009]] |
Minipreps, gel (K235030 Kan, K235030 Chl, K235031 Chl, E0840 Amp, and K115020 Amp) | Minipreps, gel (K235030 Kan, K235030 Chl, K235031 Chl, E0840 Amp, and K115020 Amp) | ||
| - | Lab Oct 13 2009 | + | [[Lab Oct 13 2009]] |
Broth cultures | Broth cultures | ||
| - | Lab Oct 14 2009 | + | [[Lab Oct 14 2009]] |
Flourescence microscopy, gel (K235032 Kan, K235033 Kan, K235036 Kan, K235037 Chl) | Flourescence microscopy, gel (K235032 Kan, K235033 Kan, K235036 Kan, K235037 Chl) | ||
| - | Lab Oct 15 2009 | + | [[Lab Oct 15 2009]] |
Flourescence microscopy,assemblies, broth cultures | Flourescence microscopy,assemblies, broth cultures | ||
| - | Lab Oct 16 2009 | + | [[Lab Oct 16 2009]] |
Latest revision as of 19:59, 17 October 2009
Current Parts
Assembly Parts
The first time a part shows up in a gel it is bold.
Preparing buffers
Making plates
Starting to make Qiagen reagents
pH'ing Tris and other stocks
More Qiagen work
Test Quiagen Miniprep and gel
Prepare CCMB80 buffer
Make SOB medium
Checking on failed DH5alpha plates
Diagnosing contamination problems
New DH5alpha plates
Acheived DH5α growth
Make competent cells following top-10 protocol
Measurement of competence
Failed competence, started on BCMB301 competent cells protocol
Test of competence on new cells, greater quantity of pUC18
Successful test of competence with pUC18
Initial messing with rehydration protocol
Successful biobrick transformation
Nandrop rehydrations, more transformations
Failed lambda cI promoter
Checked weekend culture of RFP
Another nanodrop, more rehydrations and transformations, ran a gel (B0015 Amp/Kan, J06504 Amp, R0051 Amp, B0034 Amp)
More transformations, struggling with I0500
Struggling with Kan growth, more nanodrop rehydrations
Tried to induce high copy for I0500 using IPTG
Make ccdB competent cells, miniprep plasmids, more transformations
Try again at making ccdb strains with Kan and Cm resistances, more transformations
Check on ccdB strains, minipreps, etc.
Make new ccdB competent cells, check on cultures, miniprep, gels (P1010 Kan, J23102, J45120, J45200, K145303, P0151, I0500, C0083, E0422, R1051)
Brainstorming, trying to transform P1010
Possible success in transforming P1010
Miniprep and check plasmids, cut and ligated and transformed first assemblies, a few broth cultures, gel (P1010 Kan, P1010 Cm C)
Transformed second assemblies, gel (digests)
Gels (K235000, K235001 and K235002)
Gel (linearized DNA)
Gels and assemblies (K145303 better)
Pour Cm plates
Re-plate ligation products, new competent cells, failed 2% gels (digests)
Re-plate remaining assemblies, start fridge and freezer cleanup, gels (digests)
Brainstorming
Plan for the week, gels (?)
Check P1010Kan, broth culture, gel (K081005, K098988)
Reassembling K235003, 05 and 10, starting J04450, plating stabs, talking to many people
Broth cultures
Lab move, miniprep and gels (K235003 Cm, K235005 Cm, K235010 Cm, J04450 Kan (+IPTG), J04450 Kan (-IPTG), P1010 Kan, P1010 Amp, R0051 Amp, I0500 Kan (+IPTG), I0500 Kan (-IPTG))
Scheduling a lab meetup for Wednesday, some assemblies, a subculture
Talk-talk and resources
Transformations
Broth cultures and rehydrations
Minipreps and broth cultures
Aborted miniprep
Broth cultures
Minipreps
Gels (K235009, C0012, K235013, K115017, K235014, E0240, K235016, K235017, R0051)
Flourescence microscopy - failed
Assemblies, transformations
Broth cultures not done
Broth cultures
Miniprep, gels (failed)
Subcultures
Check cultures
Transformations
Broth cultures and transformations
Minipreps and gel (K235019 Amp, K235020 Amp, K235021 Amp, K115017 Amp, C0012 Amp, J04630 Amp, J23070 Amp, R0051 Amp)
Minipreps
Broth cultures
Minipreps, gel (K235030 Kan, K235030 Chl, K235031 Chl, E0840 Amp, and K115020 Amp)
Broth cultures
Flourescence microscopy, gel (K235032 Kan, K235033 Kan, K235036 Kan, K235037 Chl)
Flourescence microscopy,assemblies, broth cultures
