TG1 is derived from E. coli K-12 strain, which is the fastest growing clone of Escherichia coli at present. At 37 ℃ on the plate, 7h can be cloned. The main strain of phage display, but also can be used to construct plasmid, lacIqZ Delta M15 which can be used for the blue white screening experiments; but without nuclease endA1 mutation, high in vivo nucleic acid enzyme content, using plasmid extraction kit to protein solution to removing nuclease in vivo of extraction plasmids. The TG1 strain needs to be cultured in the LB without antibiotics at ℃, and then 20% of the glycerol is used to preserve the bacteria.
Introduction
Alternative name TG1 Escherichia coli Strains,TG1,Escherichia coli
Strains Resistance None of resistance
Culture Medium LB
Condition 37℃ ,under the aerobic conditions
Genotype
[F´tra D36proABlacIqZΔM15]supEthi-1Δ(lac-proAB)Δ(mcrB-hsdSM)5(rK–mK–)
Minimal Medium plate
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.
