Dr. David W. Andrews

RECIPES

> DNA

> CB 10X T4 DNA Ligase (+ATP)

> Plasmid Preps

> TE (Tris-EDTA Buffer)

> Buffering Phenol

> Enzyme Buffers

> DNA gel loading buffer (5 X)

> (TBE) Tris-Borate

> TBE DNA gel running buffer (Tris borate EDTA) 10 X

> Tris-Acetate (TAE)

> Transcription Recipes

Plasmid Preps

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Solution I (Plasmid Prep)	Glucose Buffer
	100mL	500mL
0.05M glucose (m.w. 180.16)	0.9g	4.504g
0.025M Tris (pH 8.0)	1.25mL	6.25mL of (2M stock) 0.01M EDTA
	2 mL	10mL of (0.5M stock)

pH 8.0 (should be) adjust if not

SOLUTION II (MAKE FRESH)

1% SDS	5mL of 20% SDS stock
0.2N NaOH	5mL of 4N NaOH stock
	90 mL dH₂O

Note: put the water in container first as NaOH + SDS will ppt!

SOLUTION III
	1000mL	500mL	250mL
3.0M KoAC	294g	147g	73.5g
2.0M HoAC	115mL	57.5mL	28.75mL
Make up to required volume with dH₂O

pH 4.8

Keep at -20 C! but do not let freeze.

I don't really know why this solution does not keep at room temperature - but it does not.

If the freezer gets too cold you can freeze solution III but that tends to separate the solution so that you end up with frozen water on the top. This is caused by repeated freezing and thawing. It thaws during the day when people open the freezer door often and then re-freezes overnight. If you mix it really well before use it should be fine.

PEG IN NaCl
13% PEG 8000 13 g
1.6 M NaCl 40 mL 4 M NaCl

Make up to 100 ml with d H₂O