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Team:Stanford/Notebook/Lab Work/Week 9
From 2010.igem.org
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== 8/23 Monday == | == 8/23 Monday == | ||
| + | ===Chris's Notebook=== | ||
| + | |||
| + | Miniprep E1 for testing with the plate reader | ||
| + | Miniprep of PknH+B0034, Mstp+B0034; sent to sequencing | ||
| + | |||
| + | Inoculations | ||
| + | E1, PknH, MstP, EmbR (for freezer stock) | ||
| + | |||
| + | Restriction Digest | ||
| + | 4K5 and 4A5 (w/ccdb), I0500, Mstp+B0034, PknH+B0034 | ||
| + | |||
| + | Ligation | ||
| + | Mstp+B0034: 1600 bp (320 ng) | ||
| + | PknH+B0034: 1300 bp (260 ng) | ||
| + | EmbR: 1200 bp (260 ng) | ||
| + | F2620: 1060 bp (220 ng) | ||
| + | I0500: 1200 bp ( 220 ng) | ||
| + | J23100+B0034: 100 bp (0.5 ul) | ||
| + | pSB4A5: 150 ng (3400 bp) | ||
| + | pSB4K5: 150 ng (3400 bp) | ||
| + | |||
| + | I0500+Mstp+B0034 AMP | ||
| + | I0500+PknH+B0034 AMP | ||
| + | F2620+Mstp+B0034 KAN | ||
| + | F2620+PknH+B0034 KAN | ||
| + | J23100+B0034+EmbR C | ||
== 8/24 Tuesday == | == 8/24 Tuesday == | ||
| + | ===Chris's Notebook=== | ||
| + | |||
| + | Sequencing Results: Will use MB3 and PB1; both came back good | ||
| + | |||
| + | Restriction Digests: | ||
| + | MB3, PB1; use Ryan's protocol | ||
| + | Freezer Stocks | ||
| + | E1, PknH, MstP, EmbR | ||
| + | Ligations | ||
| + | I0500+Mstp+B0034 AMP | ||
| + | I0500+PknH+B0034 AMP | ||
| + | F2620+Mstp+B0034 KAN | ||
| + | F2620+PknH+B0034 KAN | ||
| + | J23100+B0034+EmbR C | ||
== 8/25 Wednesday == | == 8/25 Wednesday == | ||
| + | ===Chris's Notebook=== | ||
| + | |||
| + | Miniprep | ||
| + | MB3, I0500 (stimulate with IPTG at 1030; inoculated at 1130 pm last night) | ||
| + | Restriction Digest | ||
| + | MB3, I0500 (use Ryan's protocol; do in group of two and concentrate) | ||
| + | Ligations of Yesterday | ||
| + | F2620+PknH+B0034 KAN worked | ||
| + | J23100+B0034+EmbR AMP worked | ||
| + | I0500+PknH+B0034 AMP no growth | ||
| + | |||
| + | Ligations Today | ||
| + | |||
| + | I0500+PknH+B0034 AMP no growth (use my I0500)3 | ||
| + | I0500+Mstp+B0034 AMP | ||
| + | F2620+Mstp+B0034 KAN | ||
| + | |||
| + | Inoculate/for Testing | ||
| + | E1, MB3 | ||
== 8/26 Thursday == | == 8/26 Thursday == | ||
| + | ===Chris's Notebook=== | ||
| + | |||
| + | Transformations | ||
| + | I0500+PknH+B0034 AMP (Alex's I0500 and Jerome's I0500) | ||
| + | Miniprep | ||
| + | MB3 | ||
| + | Restriction Digest | ||
| + | MB3 (X/P) | ||
| + | Ligations | ||
| + | I0500+Mstp+B0034 AMP | ||
| + | F2620+Mstp+B0034 KAN | ||
| + | |||
| + | Inoculate (in at 1230 AM) | ||
| + | |||
| + | 3 each from successful transformations (12 total) | ||
== 8/27 Friday == | == 8/27 Friday == | ||
| + | ===Chris's Notebook=== | ||
| + | |||
| + | Ligations | ||
| + | I0500+Mstp+B0034 AMP | ||
| + | F2620+Mstp+B0034 KAN | ||
| + | |||
| + | Transform (see above) | ||
| + | |||
| + | Restriction Digest Screen | ||
| + | |||
| + | Miniprep: J23100+B0034+EmbR, F2620+B0034+PknH | ||
| + | |||
| + | Restriction Digest of the 12 inoculations (save some for sequencing; just get 500 ng) | ||
| + | |||
| + | Restriction Digest of J64100, psb4A5 (ccdb markers) | ||
| + | |||
| + | Inoculate: I0500+B0034+PknH | ||
</div> | </div> | ||
Latest revision as of 01:06, 27 October 2010
| Home | Project | Applications | Modeling | Parts | Team | Notebook |
Spring: Brainstorming | Spring Meetings
Summer: Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Summaries
|
8/23 Monday
Chris's Notebook
Miniprep E1 for testing with the plate reader Miniprep of PknH+B0034, Mstp+B0034; sent to sequencing
Inoculations E1, PknH, MstP, EmbR (for freezer stock)
Restriction Digest 4K5 and 4A5 (w/ccdb), I0500, Mstp+B0034, PknH+B0034
Ligation Mstp+B0034: 1600 bp (320 ng) PknH+B0034: 1300 bp (260 ng) EmbR: 1200 bp (260 ng) F2620: 1060 bp (220 ng) I0500: 1200 bp ( 220 ng) J23100+B0034: 100 bp (0.5 ul) pSB4A5: 150 ng (3400 bp) pSB4K5: 150 ng (3400 bp)
I0500+Mstp+B0034 AMP I0500+PknH+B0034 AMP F2620+Mstp+B0034 KAN F2620+PknH+B0034 KAN J23100+B0034+EmbR C
8/24 Tuesday
Chris's Notebook
Sequencing Results: Will use MB3 and PB1; both came back good
Restriction Digests: MB3, PB1; use Ryan's protocol Freezer Stocks E1, PknH, MstP, EmbR Ligations I0500+Mstp+B0034 AMP I0500+PknH+B0034 AMP F2620+Mstp+B0034 KAN F2620+PknH+B0034 KAN J23100+B0034+EmbR C
8/25 Wednesday
Chris's Notebook
Miniprep MB3, I0500 (stimulate with IPTG at 1030; inoculated at 1130 pm last night) Restriction Digest MB3, I0500 (use Ryan's protocol; do in group of two and concentrate) Ligations of Yesterday F2620+PknH+B0034 KAN worked J23100+B0034+EmbR AMP worked I0500+PknH+B0034 AMP no growth
Ligations Today
I0500+PknH+B0034 AMP no growth (use my I0500)3 I0500+Mstp+B0034 AMP F2620+Mstp+B0034 KAN
Inoculate/for Testing E1, MB3
8/26 Thursday
Chris's Notebook
Transformations I0500+PknH+B0034 AMP (Alex's I0500 and Jerome's I0500) Miniprep MB3 Restriction Digest MB3 (X/P) Ligations I0500+Mstp+B0034 AMP F2620+Mstp+B0034 KAN
Inoculate (in at 1230 AM)
3 each from successful transformations (12 total)
8/27 Friday
Chris's Notebook
Ligations I0500+Mstp+B0034 AMP F2620+Mstp+B0034 KAN
Transform (see above)
Restriction Digest Screen
Miniprep: J23100+B0034+EmbR, F2620+B0034+PknH
Restriction Digest of the 12 inoculations (save some for sequencing; just get 500 ng)
Restriction Digest of J64100, psb4A5 (ccdb markers)
Inoculate: I0500+B0034+PknH
