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

azi, E. c$ q,tte CMtat ot H1 Gffi Iran*dpt.n
The plastic stips s€rue as a bafiier 10 pr€vs liqud i.om llowlng
direcrly t om rhe r€servoi. ro lhe pap6r rowets, which witt be ptac€d on
Presoakod zera prob€ membrane was ctn at one corner to mark the
orlentation ofthe gsl, and was placed ovortheg€t, Slbsequen y, thr€€
presoaked sheets w€re placed on top ol zeta probe rnombrane, and
each lmo an bubbl€s wer6 removad afi€r €v6ry addiljon of sheet. A
slack ot soft papsr tssues (one to rwo boxes or faciar rissues,
KleenexrM) was precisely pLac€d on top oi the iihor sheols (A straight
stack of lissues is imporlant as it pr46nts papBr lrom hanging ove/ the
edges of lhe gei and conract rhe transter blftsr. This could calse the
inefiioien! lranster of ONA to tho membrane). On6 pack oI paper row€ts
were placed an the paperltssles and th€n agtass ptate a.d a medium
weight object (e.9., lead viat, gtass bor e) was ptaced on top or the
entre slack lo leep pressurg on rhe blffsr absorbing pap€r lissugs
The exposed edges ol the larg€ glass dish were covered with ptast'c toil
(saran Wap) to avoid evaporation of ths 10 X SSC burier, and ONA
transfer was conrinued lor m lo 24 hours. Th€ zsra proo€ memorane
wascareulry remov€d lrom th6 gel and p ac,.d i. n UV chamber ior one
mint e to ix th6 rranslerred DNA on the membrane. Then the
membran€ was placed in a s€al€d ptaslic baq (sea-a-mea )to ke€p th€
membrano hoist during srorag€ at 4.C.