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gclle was probed uirh rlie PCll ti.ng~iic~lt 01'700 bp ,111ii Bi('r~,l:lb gclii' \\,;is probed with
908 bp fiagmenr I'CK '1fnii.111. Tlic prube \\;IS lnbclcd w~rh tlie co~~i~ncrci;~lIy available
:\I~PII~~s' i11rcc 1'1bel111g AII pro\ ~ddd b! :\~i~c~\li~i~ii (L!Sj\i
Hc,\ii.ir~iioi~ o/ go~uitlil /),\',I
ciiiil c~ii~c~o~ripIir~i~!~\i\
I
Tlie ge1iu1111c DNA \\;I> ~ II~c~Ic~
\\IIII LIII,IL)IC
I~!,I~~CII~II c11/y1iic
;I, rollo\\b:
!U 111. cliru~iiosu~ii.~l L)NA (15-20 11g1
5 IL IOX rcalncnon bul'l'cr
5 111. rcblr1c11011 cll/~lllc (50 \lllll>i
1'11~ lirl~il vo!~i~ilt 111;1dc ilp to 50 ~IL \vi\l> htcr~le \\~ttr ;IIKI\\;I> i~ic~~bt~tcd l'or
211 nr 37 C.
2. 2 ILL oi' gel loud~ng but'li.r \.,IS ddcd lo eocii rotriciet! DNA belbre
clccrropliorc~~s Tlic rc~tricictl DNA \\;I> ~i/c-li;~ct~u~i.ircci ill O Xu" ;~g.~rusc gcl prcparcd III
IX TDE buffer nlollg \\111i
5 111, pl.~s~iiitl DNA rc>rr~crcd ~1111 I:colll ;IS \I;i~ld;ird 11i:irkcr
;III~ IS clcc~~opliu~c~ed o\e~-n~gIi~ ;I( 30 volt\ III IN TDI: bufir
3. Tlie gels wcrc i1;1111cd tiur 20 iil~ll UIIII
~lli~iiii~~~i br~ili~ilc ;it 1 11g;riiI bull;.r
and tlie
rcbtr~crcd DNA ii.ngmcnt b:inds wcre v~suali/cd on ;I UV I~o~is~ll~~i~ii~i~lur
:illd plio~oglaphcd.
Prui r,\.siiig titi!/ i rii~~~iltrri hloiiiiiq o/ ihc g1.i
1 Tlic sue fi.ecrioriatcd gcrio~il~c DNA In !lie gel bvi15 covered LII~~I 25U n~bl 11CI and
nglrated ~lii~il lie b~~oiiiupl~c~i~~l blue lurlib lo ycllo~v color (5-15 n~inutcs) tu depurintilc
DNA.
2. The gel was asslicd three ~inici w~rli dcnll~iefi~lized ukircr.
3. Tlic gel was ~ncubated w~th dcnzirurarion solur~o~i (l0X gel volumc. I .5M NaCl + 0.5 M
NaOH) for 15 miiiiltes twlcc, c;ich at room ~ciiipcraturc on a sliaker