Team:UT-Tokyo/Notebook

(Difference between revisions)

Revision as of 07:01, 19 August 2010

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

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 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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