Kuperwasser Lab | Solutions and Reagents

Stripping Buffer

1M Tris pH 6.8	6.25ml
10% SDS	20ml

Bring volume to a total of 100ml.
Add 50µl BME before use
Soak membrane for 30min at 50^oC while shaking in stripping buffer
Wash membrane 2 X 10 min in TTBS (Tris +Tween)
Continue with block and then primary antibody.
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Crystal Violet (Magic Solution)

DNA Loading Dye (for plasmid DNA)

SDS Sample Buffer (6X) Protein

Protein Lysis Buffer

	[Stock]	for 50ml	for 200ml
50mM Tris pH 7.5	1M	2.5ml	10ml
150mM NaCl	5M	1.5ml	6ml
0.5% NP40	10%	2.5ml	10ml

10X PBS (Phosphate Buffered Saline)

10X PPS (Stock Phosphate Buffer)

0.5 M EDTA

5M NaCl

100 ml	500 ml
NaCl 29.22 g	NaCl 146.1 g
Distilled water to 100 ml	Distilled water to 500 ml
Autoclave	Autoclave

10X TBE

	1 L	3 L
Tris base	108 g	324 g
Boric acid	55 g	165 g
0.5 M EDTA, pH 8.0	40 ml	120 ml

Equilibrating Phenol

1 M Tris (pH 7.5)

10X TBS (pH 7.5)

TTBS (pH 7.5)

	100 ml	500 ml
1 M Tris, pH 7.5	5 ml	25 ml
NaCl	0.9 g	4.5 g
10% Tween 20	1 ml	5 ml

TE (pH 8.0)

	100 ml	500 ml	Final conc.
1 M Tris, pH 7.5	1 ml	5 ml	0.01 M
0.5 M EDTA, pH 8.0	200 µl	1 ml	0.001 M

20 X SSC

	1 Liter	2 Liters
NaCl	175.3 g	350.6 g
Na Citrate	88.2 g	176.4 g

10 X MOPS Running Buffer

10% SDS

50 X TAE

	1 Liter	500 ml
Tris Base	242 g	121 g
Glacial Acetic Acid	57.1 ml	28.6 g
Na₂ EDTA 2H₂O	37.2 g	18.6 g
Final volume	1 Liter	500 ml

10ml 100X GLB

10 X TAE-GLB

1X Protein Transfer Buffer

	6 Liters	2 Liters	1 Liter
Tris Base	18.16 g	6.05 g	3.02 g
Glycine	86.50 g	28.83 g	14.40 g
Methanol	1200 ml	400 ml	200 ml

LB - Broth

NZY Agar

	1 Liter (Bottom)	250 ml (Top)
NaCl	5 g	1.25 g
MgSO₄ 7H₂O	2 g	0.50 g
Yeast Extract	5 g	1.25 g
Casein Amino Acids	10 g	2.50 g
	Adjust pH to 7.5 with 10N NaOH	Adjust pH to 7.5 with 10N NaOH
	Bring to final volume of 1 Liter	Bring to final volume of 250 ml
Agar	15 g	—
Agarose	—	1.75 g

Autoclave. Store at room temperature.

Phage Dilution Buffer

	10 ml	100 ml
1 M Tris, pH 7.4	200 µl	2 ml
5 M NaCl	200 µl	2 ml
1 M MgSO₄ 7H₂O	1 ml	10 ml

PA 317/HyTK Packaging Cell Media

NaCl	80 g
KCl	2 g
Na₂HPO₄ (anhydrous)	11.5 g
KH₂PO₄ (monobasic)	2 g

Sodium Phosphate Monobasic	7.7 g
Sodium Phosphate Dibasic (anhydrous)	20.5 g
Distilled water	2000 ml

MOPS	83.7 g
3M NaOAc, pH 6.5	33.4 ml (13.64 g)
0.5 M EDTA, pH 8.0	20.0 ml
Water	800 ml

Glycerol	5 ml
0.5 M EDTA	20 µl
Bromophenol Blue	25 mg
XCFF	25 mg

DMEM	400 ml
FBS	40 ml
7.5% NaHCO₃	8 ml (depending on the percent of CO₂ in incubator)
Hygromycin-B(2.5 x 10⁵ u/ml)	160 µl

100X GLB, pH 8.0	1 ml
50X TAE	2 ml
Glycerol	5 ml
Distilled water	2 ml

Solutions and Reagents

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