Reagents

Agarose (Ultra Pure, Sigma)

Ammonium acetate (5 M)

100 mM ATP (Pharmacia) stored at -80 °C (each aliquot is used only once)

Bacculovirus produced recombinant CAF-1 4 ng/|il [5]

Creatine phosphokinase (Boehringer) 2.5 mg/ml in H2O (!)

Cytosolic extracts derived from HeLa cells [2] © (

Ethidium bromide, 10 mg/ml (GIBCO BRL)

Loading buffer 5x: 0.42% bromophenol blue, 50% glycerol

Nuclear extracts from HeLa (4C Biotech/ Belgium or [2])

aP32 dCTP, 3000 Ci/mmol (ICN)

Phenol: chloroform: isoamyl alcohol (25 : 24: 1) (GIBCO BRL)

Proteinase K, 20 mg/ml (Roche), aliquots stored at -20 °C

5x Reaction buffer: 25 mM MgCl2, 200 mM Hepes-KOH (pH 7.8), 2.5 mM dithiothreitol (DTT; Sigma), 200 mM phosphocreatine (di-Tris salt, Sigma) ©

RNase A (Roche) 10 mg/ml in 0.01 M sodium acetate (pH 5.2), heat for 15 min at 100 °C and adjust the pH with 1 vol of Tris-HCL (pH 7.4), aliquots stored at -20 °C

Stop-mix: 30 mM EDTA, 0.7% SDS TAE 50x: 242 g Tris base, 57.1 ml glacial acetic acid, 37.2 g Na2EDTA, (pH 8.0), adjust to a final volume of 1 l with deionized water (or 50x TAE from Bio Media)

UV treated Bluescript plasmid (pBS) DNA (50 ng/^l) [1]

Was this article helpful?

0 0
Anti Ageing And Skincare Made Easy

Anti Ageing And Skincare Made Easy

Not Too Happy With Your Dull, Lifeless Skin? Do You Get Self-Conscious About Your Looks Especially The Evening You Need To Attend A Party? Are You Frustrated With Your Futile Attempts To Get Back Your Once-Glowing Skin? Fret Not Finally Discover How You Can Bring In A Natural Glow To Your Skin And Uncover Super Effective Tips And Strategies To Enliven Your Looks Once Again Enjoy A Perfectly Healthy Skin That Breathes With HappinessAnd Make Every Head Turn To Take A Second Look At You

Get My Free Ebook


Post a comment