Biology Reference
In-Depth Information
Lysis buffer (150 mM NaCl; 50 mM Tris-HCl, pH 8.5)
Add 8.77 g of NaCl, 6.05 g of Tris base in 700 ml H
2
O. Titrate to
pH 8.5 with HCl and adjust volume to 1 L with ddH
2
O. Filter
sterilize and aliquot in appropriate sizes. Store at 4 °C.
5 M NaCl
Dissolve 292.2 g of NaCl in 800 ml of ddH
2
O. Stir and adjust
volume to 1 L with ddH
2
O. Filter sterilize and aliquot in
appropriate sizes.
1× NW (Nylon Wash) Buffer
Dissolve 75.8 g Na
2
HPO
4
⋅7H
2
O and 20.83 g Na
4
EDTA⋅2H
2
O in
2.5 L of H
2
O. Titrate to pH 7.2 with H
3
PO
4
. Carefully add
500 g of SDS (Pierce, cat. no. PI28312), cover the beaker with
cling film, and stir until completely dissolved (several hours to
overnight). Adjust volume to 3.58 L with ddH
2
O and aliquot
in appropriate sizes.
PEI at 1 mg/ml
Dissolve 250 mg of PEI in 200 ml of ddH
2
O. Adjust pH to 1.9 with
approximately 400 μl of concentrated HCl. Stir until PEI is com-
pletely dissolved. Adjust pH to 7.0 with concentrated NaOH.
Adjust final volume to 250 ml. Filter sterilize through a 0.45μm
nitrocellulose filter. Store at −80 °C in 750 μl aliquots.
20× SSC
Add 175.3 g of NaCl and 88.2 g of sodium citrate in 800 ml of
ddH
2
O. Adjust pH to 7.0 with HCl. Adjust volume to 1 L.
Sterilize by autoclaving.
50× TAE
Add 242 g of Tris base, 57.1 ml of glacial acetic acid, 100 ml of
0.5 M EDTA, pH 8.0 and bring volume with ddH
2
O to 1 L.
Sterilize by autoclaving.
1× TE (10 mM Tris-HCl; 1 mM EDTA)
Add 5 ml 1 M Tris, pH 8.0, 1 ml 0.5 M EDTA, pH 8.0 and bring
volume with ddH
2
O to 500 ml. Sterilize by autoclaving.
5× TD
Add 1.01 g of MgCl
2
, 0.93 g of KCl in 300 ml of ddH
2
O, and
500 ml of 10× PBS. Stir and adjust volume to 1 L with H
2
O.
Filter sterilize and aliquot in appropriate sizes. Store at 4 °C.
1× TD/1 M NaCl
Add 29.22 g of NaCl in 100 ml of 5× TD. Adjust volume to
500 ml with ddH
2
O. Filter sterilize and aliquot in appropriate
sizes. Store at 4 °C.
10× Tris-Glycine
Add 30.27 g Tris and 144.15 g glycine in 800 ml of H
2
O. Stir and
adjust volume to 1 L.
Search WWH ::
Custom Search