Template:Kyoto/Notebook/Sep 16
From 2013.igem.org
(Difference between revisions)
(→Liquid Culture) |
(→Colony PCR) |
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!Sample||base pair | !Sample||base pair | ||
|- | |- | ||
- | | | + | |pSB1C3+P-KaiBC①||283 |
+ | |- | ||
+ | |pSB1C3+P-KaiBC②||283 | ||
|} | |} | ||
{| class="wikitable" | {| class="wikitable" | ||
!PreDenature||Denature||Annealing||Extension||cycle | !PreDenature||Denature||Annealing||Extension||cycle | ||
|- | |- | ||
- | | | + | |94°C||94°C||55°C||68°C||-- |
|- | |- | ||
- | | | + | |2min||30s||30s||18s||30cycles |
|} | |} | ||
+ | |||
===Gel Extraction=== | ===Gel Extraction=== | ||
<div class="experiment"> | <div class="experiment"> |
Revision as of 06:04, 24 September 2013
Contents |
Sep 16
Liquid Culture
Sample | medium |
---|---|
pSB1C3+P-KaiBC① | Plusgrow medium(+CP) |
pSB1C3+P-KaiBC② | Plusgrow medium(+CP) |
Colony PCR
Sample | base pair |
---|---|
pSB1C3+P-KaiBC① | 283 |
pSB1C3+P-KaiBC② | 283 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
2min | 30s | 30s | 18s | 30cycles |
Gel Extraction
Lane | DNA | Enzyme |
---|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(・▽・) |
PCR
genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total |
---|---|---|---|---|---|---|---|---|
0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 25 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
°C | °C | °C | °C | -- |
(◎Д◎) |
Electrophoresis
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
(°Д°) |
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 |
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 |
PCR
genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total |
---|---|---|---|---|---|---|---|---|
0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 25 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
°C | °C | °C | °C | -- |
(◎Д◎) |