7.8

From CSBLwiki

Contents 1 PCR (alpha-Taq pol, Sdl colony boiling) 1.1 Reaction mixture 1.2 Thermal PCR condition 2 전기영동 3 MMG Agar(100ml)

PCR (alpha-Taq pol, Sdl colony boiling)

(colony boiling = colony를 깨서 다운시킨 후 soup)

Reaction mixture

10x alpha-Taq reaction buffer 5 uL

dNTP mix(2.5mM) 3 uL

primer 2 uL

templete-genomic DNA 3 uL

alpha-Taq(2.5U/ul) 0.5ul

DW up to 50ul

Thermal PCR condition

(보통 extension 0.5-1min/kb)

95'C 2min

95'C 20sec

55'C 2min

72'C 2min

72'C 5min

전기영동

alpha-Taq은 효율이 좋은 pol이기 때문에 0.5-1 ul만 넣고 전기영동해도 충분

MMG Agar(100ml)

Sdl 배지

2.3 % sea salt

0.1 % yeast extract

0.05 % Ammonium Cloride

50mM Tris-HCl, pH 7.4(최종)

up to 100ml DW

1.5g Agar

+ 0.2% glucose(Stock 20% glucose)