Page 1 \ ?^p 6r.1 CELL CYCLE CONTROL OF HUMAN H4 ...

,nE F: c.tl Cwle C@tol ol H1 Gfle lznxnptlan
Motqiats a'd M'thods
oreciDitalion and lh€ DNA was dissolv6d in I0 lo 20 pl 01 TE_butter'
In psrallellolhe abovo procedue involving the genomlc DNA samples'
2 rg ol plasmid oNA (19, the wildtvpe FOIO8 H4 promorcr CAT
rcporter gene clnstruct) was also dig€sbd as a control ln a 200 xl ot
rcaclron, rhe plasnid DNA (mncentraiion 0 01 pg//l) was incubated
with Hind lll enzyme overnighl follo$ed by heat inacilvalion ior 10
rninltes al65'c. Subsequenlly diLuiions were mad€ to Prepar€ gel
erectrophoresis samples containing exactly 10 pg 20 p9 50 pg and
1cO pg ol digested pDNA Thls slandard curue ol digested plasrnid
oNA, as well as molecular w€ight markers (Hr:jln/EcoRl digesl ol
lambda phage DNA) was Io€ded along side 10 rrg or each ol ths
senomic oNA samPles on a 1% TAE (Tis/Acetate/EDTA pH 8 0)
All genomic DNAS wera qoantiiated by speclrophoromslrv lercre
eecifophoresls 10 ensurs thd 6xacl amounts ot DNA were loaded on
the gel. The dgesled DNA safipes w€re r6solv6d by electrophoresis
whichwas performed at 50 v tor 1a hr' G€ls were stained with ethldium
b.omide in disli{ed wale. and deslained lor 30 minrites in dislilled H2o'
DNA was visuaLized by lonq wave (312 nm) UV iuorescenc€ and
phorographed w.rh Polaroid Tvp3 57 h m (se8 Fg 16)
7A