Team:UT-Tokyo/Notebook

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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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