Template:Kyoto/Notebook/Sep 9
From 2013.igem.org
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|5m||30s||30s||40s||30cycles | |5m||30s||30s||40s||30cycles | ||
+ | |} | ||
+ | </div> | ||
+ | ===Gel Extraction=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">じっけんしゃ</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||DNA||Enzyme | ||
+ | |- | ||
+ | |(>ω・)b || || | ||
+ | |- | ||
+ | | || || | ||
+ | |} | ||
+ | [[File:igku_xxbeforexx.xxx]]<br> | ||
+ | [[File:igku_xxafterxx.xxx]]<br> | ||
+ | {| class="wikitable" | ||
+ | !Name||quantity||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |(´ω`)|| || || || | ||
+ | |- | ||
+ | |(ΦωΦ^)|| || || || | ||
+ | |} | ||
+ | </div> | ||
+ | ===Ligation=== | ||
+ | |||
+ | <div class="experiment"> | ||
+ | <span class="author">じっけんしゃ</span> | ||
+ | {| class="wikitable" | ||
+ | !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | ||
+ | |- | ||
+ | |experiment||name||xx µg/mL||name||xx µg/mL||xx µg/mL | ||
+ | |- | ||
+ | |NC||name||xx µg/mL||name||xx µg/mL||xx µg/mL | ||
|} | |} | ||
</div> | </div> |
Revision as of 18:29, 23 September 2013
Contents |
Sep 9
PCR
KaiA(plasmid) | 10x buffer | dNTPs | MgSO4 | Primer(661-RBS-KaiA-fwd) | Primer(kaiA-662-rev-ver2) | Kod-plus ver2 | MilliQ | total |
---|---|---|---|---|---|---|---|---|
1.0 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.5 | 25 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 98°C | 58.5°C | 68°C | -- |
2min | 10s | 30s | 30s | 30cycles |
KaiB(plasmid) | 10x buffer | dNTPs | MgSO4 | Primer(661-RBS-KaiA-fwd-ver2) | Primer(kaiB-663-rev-ver2) | Kod-plus ver2 | MilliQ | total |
---|---|---|---|---|---|---|---|---|
1.0 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.5 | 25 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 98°C | 58.5°C | 68°C | -- |
2min | 10s | 30s | 30s | 30cycles |
P-KaiBC(PCR products) | 10x buffer | dNTPs | MgSO4 | Primer(prefix-P-kaiBC-fwd-ver2) | Primer(P-KaiBC-suffix-rev-verw) | Kod-plus ver2 | MilliQ | total |
---|---|---|---|---|---|---|---|---|
0.7 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.8 | 25 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 98°C | 58.5°C | 68°C | -- |
2min | 10s | 30s | 30s | 30cycles |
DT(plasmid) | 10x buffer | dNTPs | MgSO4 | Primer(664-DT-fwd-ver2) | Primer(DT-suffix-660-rev-ver2) | Kod-plus ver2 | MilliQ | total |
---|---|---|---|---|---|---|---|---|
1.0 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.5 | 25 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 98°C | 58.5°C | 68°C | -- |
2min | 10s | 30s | 30s | 30cycles |
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
9/8 pSB1C3 | 83.2 | 1.45 | 0.76 |
9/8 RBS-lysis2-DT | 72.8 | 1.67 | 0.97 |
9/8 spinach-DT | 152.6 | 1.88 | 1.97 |
Nuclease-Free water | -0.4 | -14.30 | 0.25 |
Restriction Enzyme Digestion
DNA | restricion enzyme | restriction enzyme | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | (°°) µL | µL | µL | µL | µL | µL | µL |
NC | µL | µL | µL | µL | µL | µL | µL |
Transformation
Name | Sample | Competent Cells | Total | Plate |
---|---|---|---|---|
いでんし | なんちゃらµL | ごにょごにょµL | ほにゃららµL | こーせーぶっしつ |
いかどうよう | ||||
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
Colony PCR
Sample | base pair |
---|---|
9/8 Plac-spinach-DT-(1) | 659 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5m | 30s | 30s | 40s | 30cycles |
Sample | base pair |
---|---|
9/8 Plac-spinach-DT-(2) | 659 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5m | 30s | 30s | 40s | 30cycles |
Sample | base pair |
---|---|
9/8 Pcon-pt181atte-DT-(1) | 584 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5m | 30s | 30s | 40s | 30cycles |
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
(>ω・)b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|---|
(´ω`) | ||||
(ΦωΦ^) |
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL |