Team:UT-Tokyo/Notebook

From 2010.igem.org

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Our experiment notebooks (Japanese) can be read [http://www.igem-ut.net/blog/  here (www.igem-ut.net/blog) ].
Our experiment notebooks (Japanese) can be read [http://www.igem-ut.net/blog/  here (www.igem-ut.net/blog) ].
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== '''Protocol''' ==
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=== Transformation ===
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-preparation
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・iGEM parts / ligation products
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・LBbroth (No antibiotic)　500uL
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・TE　15uL
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・competent cells
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・plates
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・ice box
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・heat block(42℃)
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-Notice
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competent cells
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→ always onice!!! Melt on ice! Mix DNA as soon as cells melt!
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-Protocol
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iGEMパーツを解凍する場合
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↓P200のピペットマンにチップを付け、チップでフィルムに孔を空ける。
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↓add 15µl TE（MilliQで可）、ピペッティング
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↓1µlをコンピテントセルにいれ30min　on　ice(コンピに対し1/5倍量以下になるように)
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iGEM parts
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Hole film with a tip
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↓Add 15 uL TE or MilliQ, pipetting
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↓Mix 1uL with competent cells 50uL
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↓on ice 30 min
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↓42℃ 45 sec
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↓on ice 10 min
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↓Mix LBbroth 300 uL
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↓37℃ 30 min or room temprature 15 min (amp:omittable)
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↓Plate, 37℃ incubation over night
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Ligation products
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↓Mix ligation products 10uL, competent cells 50uL
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↓on ice 30 min
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↓42℃ 45 sec
 +
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↓on ice 10 min
 +
 +
↓Mix LBbroth 300 uL
 +
 +
↓37℃ 30 min or room temprature 15 min (amp:omittable)
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↓Plate, 37℃ incubation over night
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</div>
</div>
{{UT-Tokyo_FOOTER}}
{{UT-Tokyo_FOOTER}}

Revision as of 06:59, 19 August 2010

Notebook

Our experiment notebooks (Japanese) can be read [http://www.igem-ut.net/blog/ here (www.igem-ut.net/blog) ].


Protocol

Transformation

-preparation ・iGEM parts / ligation products ・LBbroth (No antibiotic)　500uL ・TE　15uL ・competent cells ・plates ・ice box ・heat block(42℃)

-Notice competent cells → always onice!!! Melt on ice! Mix DNA as soon as cells melt!

-Protocol iGEMパーツを解凍する場合 ↓P200のピペットマンにチップを付け、チップでフィルムに孔を空ける。 ↓add 15µl TE（MilliQで可）、ピペッティング ↓1µlをコンピテントセルにいれ30min　on　ice(コンピに対し1/5倍量以下になるように)

iGEM parts

Hole film with a tip

↓Add 15 uL TE or MilliQ, pipetting

↓Mix 1uL with competent cells 50uL

↓on ice 30 min

↓42℃ 45 sec

↓on ice 10 min

↓Mix LBbroth 300 uL

↓37℃ 30 min or room temprature 15 min (amp:omittable)

↓Plate, 37℃ incubation over night



Ligation products

↓Mix ligation products 10uL, competent cells 50uL

↓on ice 30 min

↓42℃ 45 sec

↓on ice 10 min

↓Mix LBbroth 300 uL

↓37℃ 30 min or room temprature 15 min (amp:omittable)

↓Plate, 37℃ incubation over night


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