ࡱ> +-*g i bjbjVV *r<r<ixx         $4 //Dv  :,ud. Z0 "  (0 "+G//" x : KRISHAN STAIN FOR DNA CELL CYCLE ANALYSIS Prepare a single cell suspension with .1 to 1 x 10-6 cells per tube. Wash cells with PBS, then centrifuge to pellet them. Remove the supernatent PBS, vortex to dislodge the pellet, and add 1 ml of Krishans stain. Vortex again. Refrigerate and let stand at least overnight before analysis. KRISHAN STAIN Reference: Krishan, A. (1975) J. Cell Biol.; 66,188. Stain: 0.224 g NaCit.2H2O 9.22 mg Propidium Iodide (carcinogen) ml 1% NP40 in H2O ml 1mg/ml Rnase, (DNase free) qs to 200 ml with H2O Storage: refrigerate Shelf life: at least 6 months Flow Cytometric Analysis Cells are analyzed using a Beckman Coulter FC500 flow cytometer. Doublets are excluded from the analysis using the peak vs. integral gating method. ModFit LT software (Verity Software House, Topsham, ME) is used for cell cycle analysis.   i hWShh{g*+pqO P ^ _   ( * @ _ ` a z { i ^ & F & F & F$a$(/ =!"#$% ^ 2 0@P`p2( 0@P`p 0@P`p 0@P`p 0@P`p 0@P`p 0@P`p8XV~_HmH nH sH tH 8`8 Normal_HmH sH tH DA D Default Paragraph FontViV  Table Normal :V 44 la (k (No List PK![Content_Types].xmlN0EH-J@%ǎǢ|ș$زULTB l,3;rØJB+$G]7O٭V$ !)O^rC$y@/yH*񄴽)޵߻UDb`}"qۋJחX^)I`nEp)liV[]1M<OP6r=zgbIguSebORD۫qu gZo~ٺlAplxpT0+[}`jzAV2Fi@qv֬5\|ʜ̭NleXdsjcs7f W+Ն7`g ȘJj|h(KD- dXiJ؇(x$( :;˹! I_TS 1?E??ZBΪmU/?~xY'y5g&΋/ɋ>GMGeD3Vq%'#q$8K)fw9:ĵ x}rxwr:\TZaG*y8IjbRc|XŻǿI u3KGnD1NIBs RuK>V.EL+M2#'fi ~V vl{u8zH *:(W☕ ~JTe\O*tHGHY}KNP*ݾ˦TѼ9/#A7qZ$*c?qUnwN%Oi4 =3N)cbJ uV4(Tn 7_?m-ٛ{UBwznʜ"Z xJZp; {/<P;,)''KQk5qpN8KGbe Sd̛\17 pa>SR! 3K4'+rzQ TTIIvt]Kc⫲K#v5+|D~O@%\w_nN[L9KqgVhn R!y+Un;*&/HrT >>\ t=.Tġ S; Z~!P9giCڧ!# B,;X=ۻ,I2UWV9$lk=Aj;{AP79|s*Y;̠[MCۿhf]o{oY=1kyVV5E8Vk+֜\80X4D)!!?*|fv u"xA@T_q64)kڬuV7 t '%;i9s9x,ڎ-45xd8?ǘd/Y|t &LILJ`& -Gt/PK! ѐ'theme/theme/_rels/themeManager.xml.relsM 0wooӺ&݈Э5 6?$Q ,.aic21h:qm@RN;d`o7gK(M&$R(.1r'JЊT8V"AȻHu}|$b{P8g/]QAsم(#L[PK-![Content_Types].xmlPK-!֧6 0_rels/.relsPK-!kytheme/theme/themeManager.xmlPK-!0C)theme/theme/theme1.xmlPK-! ѐ' theme/theme/_rels/themeManager.xml.relsPK] i i i 8@0(  B S  ?;;DkBFFk9*urn:schemas-microsoft-com:office:smarttagsState8*urn:schemas-microsoft-com:office:smarttagsCity9*urn:schemas-microsoft-com:office:smarttagsplace  ljq fpkk33{kkyR\DI\^6n''^'`o(.0^`o(./ 0/ ^/ `0o(..0^`0o(... 0^`0o( .... ^`o( ..... ^`o( ...... `^``o(.......  ` ^ ``o(........hh^h`o(.''^'`o(.N 0N ^N `0o(.. 0 ^ `0o(... 0^`0o( .... ^`o( .....   ^ `o( ...... &`&^&``o(....... +`+^+``o(........DI\^yR 0E{gWSik@iX@UnknownG* Times New Roman5Symbol3. * ArialACambria Math"qha#24hhKHP ?WS2!xx )KRISHAN STAIN FOR DNA CELL CYCLE ANALYSISHelmK Helm, Karen  Oh+'0  < H T`hpx,KRISHAN STAIN FOR DNA CELL CYCLE ANALYSISHelmKNormal Helm, Karen2Microsoft Office Word@F#@*d@*dpirationDate _SourceUrl _SharedFileIndex 1000.00000000000*KRISormDocumentLibraryFormDocumentLibraryFormffice/2006/metadata/longProperties"/> repoint/v3/contenttype/forms">DocumentLibraryFItem  Properties3154Y1JMDT==2 zzItem  #KProperties4BECCWWA==2zzItem  PropertiesOccurs="0" maxOccurs="1" ma:index="4" ma:displayName="Title"/> This value indicates the number of saves or revisions. The application is responsible for updating this value after each revision. cumentation> This value indicates the number of saves or revisions. The application is responsible for updating this value after each revision. > ds:itemID="{9B75E9FD-225C-4A04-BC0B-CCC25B397158}" xmlDocumentLibraryF՜.+,D՜.+,X hp  UCHSCh *KRISHAN STAIN FOR DNA CELL CYCLE ANALYSIS Title  `  xd_SignatureOrder TemplateUrl xd_ProgIDPublishingStartDatePublishingEx