Prepare a DNA master mix cocktail consisting of Per Rxn For 6 Rxns DNA 25g 1250l 750l 10X NEB Sau3A I Buffer 15l Sau3A I storage buffer 10mM Tris pH 74 50mM KCl 1mM DTT 0 ID: 958591
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Pilot Sau3A Digestion Pre
pare a DNA master mix cockt
ail consisting of:
Per Rxn For 6 Rxns DNA
(~2.5g) 12.50l **** 75
.0l 10X NEB Sau3A I Buffer
1.5l Sau3A I storage bu
ffer 10mM Tris ( pH 7.4 )
50mM KCl 1mM DTT 0.1mM
EDTA 200g/ml BSA 50% glyc
erol Stock Vol Uni
ts: (0.125U) (0.25U) (0.37
5U) (0.50U) (0.625U) (0.75
U) Vol: 1.25l 2.5l 3.75l
5.0l 6.25l 7.5l 5X G
el loading buffer with dye
(Good for just about all ap
plications) 50ml 5
X Conc 80% Glycerol 47ml
75% Br Add the appropri
ate volume of enzyme to eac
h. Aliquot 14.3l DNA coc
ktail to each of the six tu
bes. Mix gently. Incu
bate at 37 o C for 1.25h.
Add 4l Loading Dye (the E
DTA and SDS stop the reacti
on). Load onto a 0.8% TE
A agarose gel. Run gel o
vernight for 500V-h (30-35V
for ~14-16h, on a standard
sized gel). Stockinger L