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
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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