ࡱ> ,.+k  bjbj .,j,j]_____sss8sp(\^^^^^^)d^_^__s>>>>>>> H0>CC>>"C_`>^^nCY %: Oligonucleotide 5 adenylation Using a 5 phosphorylated oligonucleotide and the 5 adenylation kit from NEB Set up reaction: Phosphorylated DNA Oligonucleotide 5 l (500 pmol) 10x 5 DNA Adenylation Reaction Buffer 5 l 1 mM ATP 5 l Nuclease-free Water to 45 l Mth RNA Ligase 5 l Incubate at 65C for 2 hour. Incubate 85(C for 5min Lid at 95( Dilute to 100ul with water, extract with 100l phenol:chloroform pH8 Add 10l 3M NaOAc, 1l glycogen and 250l 100% ethanol Vortex Spin 15 min top speed at 4(C Wash pellet with 400l 70% ethanol Dissolve pellet in 50l 0.1xTE (to 10pM/l final concentration) Vortex then let sit at room temperature for 30min or so Store at -30(C      FILENAME Oligo adenylation and purification v1.1 Houseley lab  PAGE 1  no ) * @ G H S T _ i j k l {   2 3 H I s ޾޷޳hejheU h^TZh^TZh) hQh%hmh1x jhQh% jhQhQh hQhQhMh)5hM5CJ(aJ(hQ5CJ(aJ(> no ) * G H _ k l   2 `gdm`gdQgdQgd)$a$gd)2 3 s     gdQ            ü h^TZh^TZh7h0JmHnHu h(0Jjh(0JUh%hl7:hpv)mHnHuh(jh(U,1h. A!"#$% x2 0@P`p2( 0@P`p 0@P`p 0@P`p 0@P`p 0@P`p 0@P`p8XV~ 0@ 0@ 0@ 0@ 0@ 0@ 0@ 0@ 0@ 0@ 0@ 0@ 0@ 0@66666_HmH nH sH tH @`@ NormalCJ_HaJmH sH tH DA`D Default Paragraph FontRiR  Table Normal4 l4a (k (No List 4@4 Mr2Header  9r 4 @4 Mr2Footer  9r .)@. Mr2 Page NumberPK![Content_Types].xmlN0EH-J@%ǎǢ|ș$زULTB l,3;rØJB+$G]7O٭V (BO)MBT.$@0H!A>풠Uc-zD[&!rX=}zC0` ި%.]Ssd--7 +fOZեrŵVœ\lji2ZGwm-3˵j7\ Uk5FҨ-:xRkcr3Ϣ+9kji9OP Et-j|#p;E=Ɖ5Z2sgF=8 K}*7c<`*HJTcB<{Jc]\ Ҡk=ti"MGfIw&9ql> $>HmPd{(6%z:"'/f7w0qBcF6f Iöi1(\}B5ҹ~Bcr6I;}mY/lIz1!) ac 1fm ƪN^I77yrJ'd$s<{uC>== Ƌ(uX=WA NC2>GK<(C,ݖm: &-8j^N܀ݑ$4:/x vTu>*ٞn{M.Ǿ0v4<1>&ⶏVn.B>1CḑOk!#;Ҍ}$pQ˙y')fY?u \$/1d8*ZI$G#d\,{uk<$:lWV j^ZơSc*+ESa1똀 k3Ģxzjv3,jZU3@jWu;z \v5i?{8&==ϘNX1?  O4׹ӧCvHa01 %xz24ĥ=m X\(7Xjg !Ӆqd? cG7.`~w*?, 2 nN*"Fz_&n &\ F:l[+%f U OO\\\_   2   :PWY_!8@0(  B S  ?AE33?@ij{ 13BHISV[\]qs  ?@ij{ 13BHISV[\]qsF