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Chromatin Immunoprecipitation Assay and PCR
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ÈÕÆÚ£º2007-01-18 01:12:17
µã»÷£º215 ÆÀÂÛ£º0
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| Chromatin Preparation Two days before prep, inoculate 5ml media (YP + 2% Raffinose). 30 C 24 hrs. Day before prep, inoculate 300 ml media (YP + 2% Raffinose) with overnight culture. For YP-Raffinose, 300 ul wt will reach OD600=0.4 in ~16 hours. Day |
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ȾɫÖÊÃâÒß³Áµí·ÖÎö
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ÈÕÆÚ£º2007-01-18 01:00:01
µã»÷£º327 ÆÀÂÛ£º0
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| ȾɫÖÊÃâÒß³Áµí·ÖÎö £¨ Chromatin Immunoprecipitation Assay, ChiP £© ÕæºËÉúÎïϸ°û״̬ÊÇÓÉÄÚÔ´ºÍÍâÔ´ÒòËØ¹²Í¬Ó°ÏìµÄ£¬ËùÓÐÐźŴ«µÝ;¾¶µÄÖյ㶼ÊÇ DNA ¡£ DNA ͨ¹ýºË µ°°× ¸´ºÏÎï×é³ÉȾɫÖÊ£¬È¾É«ÖÊÊÇ»ùÒòµ÷¿ØµÄÒ»¸öÖØÒª×÷ÓÃλµã¡£×ªÂ¼¼¤»îÒò×Ӻ͸¨ÖúÒÖÖÆÒò×ÓµÄÑо¿ |
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General Principles of Immunoprecipitation
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ÈÕÆÚ£º2007-01-18 00:48:28
µã»÷£º136 ÆÀÂÛ£º0
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| Lysis buffer. The choice of lysis buffer depends on what kind of IP you are doing. RIPA buffer gives the lowest background, but can denature some kinases. It also has the potential to disrupt protein:protein interactions. NP40 buffer (EVF) is less d |
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Immunoprecipitation and Immune Complex kinase assay
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ÈÕÆÚ£º2007-01-18 00:46:50
µã»÷£º110 ÆÀÂÛ£º0
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| 1) In the cold room, wash cells with cold "Tris" (the same stuff used for TC). Spin down the cells if you are working with suspension cells in a low speed swinging bucket centrifuge, resuspend in approximately 1 ml Tris and spin at 2K in a microfuge |
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Chromatin Immunoprecipitation from Yeast Whole Cell Extracts
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ÈÕÆÚ£º2007-01-18 00:43:59
µã»÷£º116 ÆÀÂÛ£º0
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| 1. Crosslink protein DNA complexes in vivo Grow 50ml yeast culture to OD600 0.5-1.0. In fume hood, add 1.4ml 37% fomaldehyde solution to a 50ml conical tube. Fill tube with culture to just below the 50ml line. Incubate at room temperature for 15min. |
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ChIP Assay Protocol
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ÈÕÆÚ£º2007-01-18 00:40:03
µã»÷£º133 ÆÀÂÛ£º0
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| Assemble the following components: Cells, grown in 10 cm dishes Room Temp. PBS Cold PBS Cold Sonication Buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl pH 7.5) Prepare sonication buffer as follows: 10 ml of 10% SDS, 2 ml of 500 mM EDTA, 5 ml of 1 M Tris, |
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SouthernÔÓ½»
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ÈÕÆÚ£º2007-01-16 02:43:50
µã»÷£º192 ÆÀÂÛ£º0
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| SouthernÔÓ½»¿ÉÓÃÀ´¼ì²â¾ÏÞÖÆÐÔÄÚÇÐøÇиîºóµÄDNAƬ¶ÎÖÐÊÇ·ñ´æÔÚÓë̽ÕëͬԴµÄÐòÁÐ,Ëü°üÀ¨ÏÂÁв½Öè: (1) øÇÐDNA, Äý½ºµçÓ¾·ÖÀë¸÷øÇÐÆ¬¶Î,È»ºóʹDNAÔλ±äÐÔ¡£ (2) ½«DNAƬ¶Î×ªÒÆµ½¹ÌÌåÖ§³ÖÎï(ÏõËáÏËÎ¬ËØÂËĤ»òÄáÁúĤ)ÉÏ¡£ (3) Ô¤ÔÓ½»ÂËĤ,ÑÚ¸ÇÂËĤÉÏ·ÇÌØÒìÐÔλµã¡£ (4 |
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ÈÕÆÚ£º2007-01-15 22:56:28
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| ÎâÕñÁú Äô¾ü ±«»ÛÄþ Ðì±ø Íõºì ÖлªÎ¢ÉúÎïѧºÍÃâÒßѧÔÓÖ¾ 2000ÄêµÚ4ÆÚµÚ20¾í ϸ¾úѧ ¸±ÈÜѪÐÔ»¡¾ú(Vibrio parahaemolyticus,Vp)ÊÇÒ»ÖÖÄÜÒýÆðʳÎïÖж¾ºÍÂÃÓÎÕ߸¹ÐºµÄÊÈÑÎÐÔϸ¾ú¡£Ö±½ÓÈÜѪ¶¾ËØ(thermostable direct hemolysin,TDH)ÊÇÆäÖ÷ÒªµÄÖ²¡Òò×Ó¡£±¾Ñо¿Í¨¹ý¹¹½¨tdh |
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ÈÕÆÚ£º2007-01-14 01:31:26
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ÈÕÆÚ£º2007-01-14 01:23:37
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ÈÕÆÚ£º2007-01-14 00:49:32
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ÈÕÆÚ£º2007-01-14 00:45:34
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ÈÕÆÚ£º2006-12-16 08:57:06
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