Template:Kyoto/Notebook/Aug 10
From 2013.igem.org
(Difference between revisions)
(→Ligation) |
(→LB Medium Plate(+Ampicillin)) |
||
(35 intermediate revisions not shown) | |||
Line 1: | Line 1: | ||
==Aug 10== | ==Aug 10== | ||
- | === | + | ===Colony PCR=== |
<div class="experiment"> | <div class="experiment"> | ||
- | <span class="author"> | + | <span class="author">Nakamoto</span> |
{| class="wikitable" | {| class="wikitable" | ||
- | ! | + | !Sample |
|- | |- | ||
- | | | + | |8/9 BBa_J44000 -(1) |
|- | |- | ||
- | |NC|| | + | |8/9 BBa_J44000 -(2) |
+ | |- | ||
+ | |NC | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94 °C||94 °C||55 °C||68 °C||-- | ||
+ | |- | ||
+ | |5 min||30 sec||30 sec||30 sec||30 cycles | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Sample | ||
+ | |- | ||
+ | |8/9 BBa_J23106 | ||
+ | |- | ||
+ | |8/9 BBa_J23100 -(1) | ||
+ | |- | ||
+ | |8/9 BBa_J23100 -(2) | ||
+ | |- | ||
+ | |NC | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94 °C||94 °C||55 °C||68 °C||-- | ||
+ | |- | ||
+ | |5 min||30 sec||30 sec||1 min||30 cycles | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===Electrophoresis=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Tatsui and Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||Sample | ||
+ | |- | ||
+ | | ||100bp ladder | ||
+ | |- | ||
+ | |4||8/9 BBa_J44000 -1 | ||
+ | |- | ||
+ | |5||8/9 BBa_J44000 -2 | ||
+ | |- | ||
+ | |6||NC | ||
+ | |} | ||
+ | [[File:Igku Aug10 Electrophoresis BBa 1.jpg]]<br> | ||
+ | {| class="wikitable" | ||
+ | !Lane||Sample | ||
+ | |- | ||
+ | | ||1kbp ladder | ||
+ | |- | ||
+ | |1||8/9 BBa_J23106 | ||
+ | |- | ||
+ | |2||8/9 BBa_J23100 -1 | ||
+ | |- | ||
+ | |3||8/9 BBa_J23100 -2 | ||
+ | |- | ||
+ | |4||NC | ||
+ | |} | ||
+ | [[File:Igku Aug10electrophoresis1.jpg]]<br> | ||
+ | </div> | ||
+ | |||
+ | ===Liquid Culture=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||medium | ||
+ | |- | ||
+ | |8/9 BBa-J23106||LB(+Amp) | ||
+ | |- | ||
+ | |8/9 BBa-J23100-1||LB(+Amp) | ||
+ | |- | ||
+ | |8/9 BBa-J23100-2||LB(+Amp) | ||
+ | |- | ||
+ | |8/9 BBa-J44000-1||LB(+Amp) | ||
+ | |- | ||
+ | |8/9 BBa-J44000-2||LB(+Amp) | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===LB Medium Plate(+Ampicillin)=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto & Tatsui</span> | ||
+ | {| class="wikitable" | ||
+ | !volume||200ml | ||
+ | |- | ||
+ | |Bacto T2ypton || 2g | ||
+ | |- | ||
+ | |Bacto yeast extract ||1g | ||
+ | |- | ||
+ | |NaCl||1g | ||
+ | |- | ||
+ | |0.4M NaOHaq||500µL | ||
+ | |- | ||
+ | |Agarose Pouder||2g | ||
+ | |- | ||
+ | |Ampicillin||40µL | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===Transformation=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Hirano</span> | ||
+ | {| class="wikitable" | ||
+ | !Name||Sample||Competent Cells||Total||Plate | ||
+ | |- | ||
+ | |tetR||3µL||19µL||22µL||Kan | ||
+ | |- | ||
+ | |RBS||3µL||19µL||22µL||Amp | ||
+ | |- | ||
+ | |lacZα||3µL||19µL||22µL||CP | ||
+ | |- | ||
+ | |lacI||3µL||19µL||22µL||CP | ||
+ | |- | ||
+ | |Plac||3µL||19µL||22µL||CP | ||
|} | |} | ||
</div> | </div> |
Latest revision as of 03:40, 28 September 2013
Contents |
Aug 10
Colony PCR
Sample |
---|
8/9 BBa_J44000 -(1) |
8/9 BBa_J44000 -(2) |
NC |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94 °C | 94 °C | 55 °C | 68 °C | -- |
5 min | 30 sec | 30 sec | 30 sec | 30 cycles |
Sample |
---|
8/9 BBa_J23106 |
8/9 BBa_J23100 -(1) |
8/9 BBa_J23100 -(2) |
NC |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94 °C | 94 °C | 55 °C | 68 °C | -- |
5 min | 30 sec | 30 sec | 1 min | 30 cycles |
Electrophoresis
Lane | Sample |
---|---|
100bp ladder | |
4 | 8/9 BBa_J44000 -1 |
5 | 8/9 BBa_J44000 -2 |
6 | NC |
Lane | Sample |
---|---|
1kbp ladder | |
1 | 8/9 BBa_J23106 |
2 | 8/9 BBa_J23100 -1 |
3 | 8/9 BBa_J23100 -2 |
4 | NC |
Liquid Culture
Sample | medium |
---|---|
8/9 BBa-J23106 | LB(+Amp) |
8/9 BBa-J23100-1 | LB(+Amp) |
8/9 BBa-J23100-2 | LB(+Amp) |
8/9 BBa-J44000-1 | LB(+Amp) |
8/9 BBa-J44000-2 | LB(+Amp) |
LB Medium Plate(+Ampicillin)
volume | 200ml |
---|---|
Bacto T2ypton | 2g |
Bacto yeast extract | 1g |
NaCl | 1g |
0.4M NaOHaq | 500µL |
Agarose Pouder | 2g |
Ampicillin | 40µL |
Transformation
Name | Sample | Competent Cells | Total | Plate |
---|---|---|---|---|
tetR | 3µL | 19µL | 22µL | Kan |
RBS | 3µL | 19µL | 22µL | Amp |
lacZα | 3µL | 19µL | 22µL | CP |
lacI | 3µL | 19µL | 22µL | CP |
Plac | 3µL | 19µL | 22µL | CP |