DNA实验

Restriction Enzyme Buffer

2024-10-23 DNA实验加入收藏

Restriction Enzyme Buffer Most enzymes can use REact buffers; however, some are

Restriction Enzyme Buffer Most enzymes can use REact buffers; however, some are made up separately. Use fresh Milli-Q water , siliconized or sterile glassware or disposable plastic ware to make the following stock solutions up fresh (discard after use), combine to make 5 ml of 10X reaction buffers, put 1 ml in to each of 5 microfuge tubes (MFTs) and store the current one at -20℃ and the rest at -70℃.

A: 1 M Tris-HCl 50 ml: 4.88 g Tris-HCl + 2.3 g Tris-Base (pH 7.9 at 25℃)

B: 1 M Tris-HCl 50 ml: 6.61 g Tris-HCl + 0.8 g Tris-Base (pH 7.4 at 25℃)

C: 1 M MgCl2 10 ml: 2.03 g

D: 3 M NaCl 20 ml: 3.50 g

E: 2M KCl 10 ml: 1.48 g

Ban I: 0.3ml B + 0.3ml C + 0.10ml D + 0.1ml BSA + 21µl bME + 4.18ml H2O

Bgl II: 0.5ml B + 0.5ml C + 1.67ml D + 0.1ml BSA + 35µl bME + 2.20ml H2O

Dra I: 0.5ml A + 0.5ml C + 0.83ml D + 0.1ml BSA + + 3.06ml H2O

Hpa II: 0.5ml B + 0.5ml C + 0.25ml E + 0.1ml BSA + 35µl bME + 3.62ml H2O

Nae I: 0.5ml A + 0.5ml C + 0.33ml D + 0.1ml BSA + 18µl bME + 3.55ml H2O

Not I: 0.5ml A + 0.5ml C + 2.50ml D + 0.1ml BSA + 5µl TX100+ 1.40ml H2O

Pst I: 0.5ml B + 0.5ml C + 1.67ml D + 0.1ml BSA + + 2.23ml H2O