Template:Kyoto/Notebook/Sep 16
From 2013.igem.org
(Difference between revisions)
(→Colony PCR) |
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{| class="wikitable" | {| class="wikitable" | ||
!Lane||DNA||Enzyme | !Lane||DNA||Enzyme | ||
+ | |- | ||
+ | |1||1kb ladder | ||
+ | |- | ||
+ | |2||pT181 attenuator||X+P | ||
+ | |- | ||
+ | |3||pT181 attenuator||X+P | ||
+ | |- | ||
+ | |4||pT181 attenuator||X+P | ||
+ | |- | ||
+ | |6||pSB1C3||E+S | ||
+ | |- | ||
+ | |7||pSB1C3||E+S | ||
+ | |- | ||
+ | |8||pSB1C3||E+S | ||
+ | |- | ||
+ | |10||pSB4K5||X+P | ||
+ | |- | ||
+ | |11||pSB4K5||X+P | ||
+ | |- | ||
+ | |12||pSB4K5||X+P | ||
+ | |- | ||
|} | |} | ||
[[File:igku_xxbeforexx.xxx]]<br> | [[File:igku_xxbeforexx.xxx]]<br> | ||
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{| class="wikitable" | {| class="wikitable" | ||
!Name||concentration[µg/mL]||260/280||260/230 | !Name||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |pT181 attenuator(X+P)|| ||8.2||1.62||0.11 | ||
+ | |- | ||
+ | |pSB1C3(E+S)||49.5||1.48||0.18 | ||
+ | |- | ||
+ | |pSB4K5(X+P)||18.1||1.73||0.57 | ||
+ | |- | ||
+ | |DT(S+P)||70.4||1.36||1.02 | ||
+ | |- | ||
+ | |aptamer12-1R-DT(S+P)||42.0||1.82||0.47 | ||
+ | |- | ||
+ | |spinach-DT(S+P)||36.8||1.79||0.61 | ||
+ | |- | ||
|} | |} | ||
+ | |||
===Miniprep=== | ===Miniprep=== | ||
<div class="experiment"> | <div class="experiment"> |
Revision as of 06:10, 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 |
---|---|---|
1 | 1kb ladder | |
2 | pT181 attenuator | X+P |
3 | pT181 attenuator | X+P |
4 | pT181 attenuator | X+P |
6 | pSB1C3 | E+S |
7 | pSB1C3 | E+S |
8 | pSB1C3 | E+S |
10 | pSB4K5 | X+P |
11 | pSB4K5 | X+P |
12 | pSB4K5 | X+P |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 | |
---|---|---|---|---|
pT181 attenuator(X+P) | 8.2 | 1.62 | 0.11 | |
pSB1C3(E+S) | 49.5 | 1.48 | 0.18 | |
pSB4K5(X+P) | 18.1 | 1.73 | 0.57 | |
DT(S+P) | 70.4 | 1.36 | 1.02 | |
aptamer12-1R-DT(S+P) | 42.0 | 1.82 | 0.47 | |
spinach-DT(S+P) | 36.8 | 1.79 | 0.61 |
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◎) |