菌株名稱(chēng):E. coli HB2151 Cells
規(guī)格:100μL/管
狀態(tài):20%甘油保存
性質(zhì)參數(shù):K12 (lac-pro), ara, nalr, thi/F'[proAB, lacIq, lacZ M15].
建議:
1. 菌株保存在20%甘油的培養(yǎng)基中,接到菌株后,請(qǐng)?zhí)羧【涸?x YT-N平板(2xYT medium containing 100 μg/ml nalidixic acid)上劃線(xiàn),37℃過(guò)夜培養(yǎng)后,再次挑取單個(gè)菌落在Minimal Medium 平板上劃線(xiàn),37℃過(guò)夜培養(yǎng)后即可使用,以便使纖毛充分表達(dá),利于噬菌體感染;
2. 在Minimal Medium 平板上生長(zhǎng)的**,不宜在冰箱中放置過(guò)久再使用;
3. Minimal Medium plate 配制方法:
Prepare stocks of the following:
1 M MgCl2?6H2O: Dissolve 20.33 g in distilled water to a final volume of 100 ml and autoclave.
1 M CaCl2?2H2O: Dissolve 14.7 g in distilled water to a final volume of 100 ml and autoclave.
1 M thiamine hydrochloride: Dissolve 33.73 g in distilled water to a final volume of 100 ml and sterilize using a 0.22 μm filter.
20% glucose: Dissolve 20 g of D-(+)-glucose (anhydrous) in distilled water to a final volume of 100 ml and sterilize by filtration through a 0.2 μm filter. Do not autoclave.
In a 500 ml bottle, dissolve 6 g of Na2HPO4 (dibasic), 3 g of KH2PO4(monobasic) and 1 g of NH4Cl in distilled water to a final volume of 500 ml. adjust the pH to 7.4 with NaOH.
In a separate 1 liter bottle, add distilled water to 15 g of Bacto-agar to a final Volume of 500 ml. Autoclave both bottles simultaneously to sterilize. Cool both bottles to 50-60°C and combine. Add1 ml of 1 M MgCl2?6H2O, 1 ml of 1 M CaCl2?2H2O, 1 ml of 1 M thiamine hydrochloride and 5 ml of 20% glucose. Pour plates immediately.