Team:UT-Tokyo/Notebook
From 2010.igem.org
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