Znn3bq.jpeg
²é¿´: 2648  |  »Ø¸´: 7
¡¾½±Àø¡¿ ±¾Ìû±»ÆÀ¼Û5´Î£¬×÷ÕßyexuqingÔö¼Ó½ð±Ò 3.25 ¸ö
µ±Ç°Ö»ÏÔʾÂú×ãÖ¸¶¨Ìõ¼þµÄ»ØÌû£¬µã»÷ÕâÀï²é¿´±¾»°ÌâµÄËùÓлØÌû

yexuqing

гæ (ÎÄѧ̩¶·)


[×ÊÔ´] ¡¾ÆäËû¡¿ÊµÑéÊÒ³£¼ûΣÏÕÉú»¯Æ·¼°´¦Àí·½·¨ ÒÑÓÐ2È˲ÎÓë

ʵÑéÊÒ³£¼ûΣÏÕÉú»¯Æ·¼°´¦Àí·½·¨


1¡¢DMSO£¨Dimethyl sulfoxide£¬¶þ¼×»ùÑÇí¿£©

DMSO¿ÉÓÃ×÷ÒÒȲ¡¢·¼Ìþ¡¢¶þÑõ»¯Áò¼°ÆäËûÆøÌåµÄÈܼÁÒÔ¼°ëæÂÚÏËά·ÄË¿ÈܼÁ£¬ÊÇÒ»ÖÖ¼ÈÈÜÓÚË®ÓÖÈÜÓÚÓлúÈܼÁµÄ¼«ÎªÖØÒªµÄ·ÇÖÊ×Ó¼«ÐÔÈܼÁ£¬¶ÔƤ·ôÓм«Ç¿µÄÉøÍ¸ÐÔ£¬ÓÐÖúÓÚÒ©ÎïÏòÈËÌåÉøÍ¸£¬Ò²¿É×÷ũҩµÄÌí¼Ó¼Á£¬ÊÇÒ»ÖÖÊ®·ÖÖØÒªµÄ»¯Ñ§ÊÔ¼Á¡£

´ËÍ⣬DMSOÒ²ÊÇÒ»ÖÖÉøÍ¸ÐÔ±£»¤¼Á£¬Äܹ»½µµÍϸ°û±ùµã£¬¼õÉÙ±ù¾§µÄÐγɣ¬¼õÇá×ÔÓÉ»ù¶Ôϸ°ûË𺦣¬¸Ä±äÉúÎïĤ¶Ôµç½âÖÊ¡¢Ò©Îï¡¢¶¾ÎïºÍ´úл²úÎïµÄͨ͸ÐÔ¡£

µ«ÊÇÑо¿±íÃ÷£¬DMSO´æÔÚÑÏÖØµÄ¶¾ÐÔ×÷Óã¬Óëµ°°×ÖÊÊèË®¼¯ÍÅ·¢Éú×÷Ó㬵¼Öµ°°×ÖʱäÐÔ£¬¾ßÓÐѪ¹Ü¶¾ÐԺ͸ÎÉö¶¾ÐÔ¡£DMSOÊǶ¾ÐԱȽÏÇ¿µÄ¶«Î÷£¬ÓõÄʱºòÒª±ÜÃâÆä»Ó·¢£¬Òª×¼±¸1%-5%µÄ°±Ë®±¸Óã¬Æ¤·ôÕ´ÉÏÖ®ºóÒªÓôóÁ¿µÄˮϴÒÔ¼°Ï¡°±Ë®Ï´µÓ¡£×îΪ³£¼ûµÄ֢״Ϊ¶ñÐÄ¡¢Å»Í¡¢Æ¤Õî¼°ÔÚÆ¤·ô¡¢ºÍºô³öµÄÆøÌåÖз¢³ö´óËâ¡¢Ñó´Ð¡¢ÄµòÃζ¡£

ÎüÈ룺¸ß»Ó·¢Å¨¶È¿ÉÄܵ¼ÖÂÍ·Í´£¬ÔÎÑ£ºÍÕò¾²¡£

Ƥ·ô£ºÄܹ»×ÆÉËÆ¤·ô²¢Ê¹Æ¤·ôÓдÌÍ´¸Ð£¬ÈçͬËù¼ûµÄƤÕî¼°Ë®ÅÝÒ»Ñù¡£Èô¶þ¼×»ùÑÇí¿Ó뺬ˮµÄƤ·ô½Ó´¥£¬»á²úÉúÈÈ·´Ó¦¡£Òª±ÜÃâ½Ó´¥º¬Óж¾ÐÔÔ­ÁÏ»òÎïÖʵĶþ¼×»ùÑÇí¿ÈÜÒº£¬ÒòÆä¶¾ÐÔ²»ÎªÈËËùÖª£¬¶ø¶þ¼×»ùÑÇí¿È´¿ÉÄÜ»áÉøÈ뼡·ô£¬ÔÚÒ»¶¨Ìõ¼þÏ»ὫÓж¾ÎïÖÊ´úÈ뼡·ô¡£

2¡¢EB£¨Ethidium bromide£¬ä廯ÒÒ¶§£©

ä廯ÒÒ¶§ÊÇÒ»Öָ߶ÈÁéÃôµÄÓ«¹âȾɫ¼Á£¬ÓÃÓÚ¹Û²ìÇíÖ¬ÌǺ;۱ûÏ©õ£°·Äý½ºÖеÄDNA¡£ä廯ÒÒ¶§Óñê×¼302nm×ÏÍâ¹â͸ÉäÒǼ¤·¢²¢·ÅÉä³ö³ÈºìÉ«Ðźţ¬¹Û²ìÇíÖ¬ÌÇÄý½ºÖÐDNA×î³£Óõķ½·¨ÊÇÀûÓÃÓ«¹âȾÁÏä廯ÒÒ¶§½øÐÐȾɫ£¬ä廯ÒÒ¶§º¬ÓÐÒ»¸ö¿ÉÒÔǶÈëDNA¶Ñ»ý¼î»ùÖ®¼äµÄÒ»¸öÈý»·Æ½Ãæ»ùÍÅ¡£ËüÓëDNAµÄ½áºÏ¼¸ºõûÓмî»ùÐòÁÐÌØÒìÐÔ¡£ÔÚ¸ßÀë×ÓÇ¿¶ÈµÄ±¥ºÍÈÜÒºÖУ¬´óԼÿ2.5¸ö¼î»ù²åÈëÒ»¸öä廯ÒÒ¶§·Ö×Ó¡£µ±È¾ÁÏ·Ö×Ó²åÈëºó£¬ÆäÆ½Ãæ»ùÍÅÓëÂÝÐýµÄÖáÏß´¹Ö±²¢Í¨¹ý·¶µÂ»ªÁ¦ÓëÉÏϼî»ùÏ໥×÷Óá£Õâ¸ö»ùÍŵĹ̶¨Î»Öü°ÆäÓë¼î»ùµÄÃÜÇнӽü£¬µ¼ÖÂÓëDNA½áºÏµÄȾÁϳÊÏÖÓ«¹â£¬ÆäÓ«¹â²úÂʱÈÓÎÀëÈÜÒºÖÐȾÁÏÓÐËùÔö¼Ó¡£DNAÎüÊÕ254nm´¦µÄ×ÏÍâ·øÉä²¢´«µÝ¸øÈ¾ÁÏ£¬¶ø±»½áºÏµÄȾÁϱ¾ÉíÎüÊÕ302nmºÍ366nmµÄ¹â·øÉä¡£ÕâÁ½ÖÖÇé¿öÏ£¬±»ÎüÊÕµÄÄÜÁ¿Ôڿɼû¹âÆ×ºì³ÈÇøµÄ590nm´¦ÖØÐ·¢Éä³öÀ´¡£ÓÉÓÚEB-DNA¸´ºÏÎïµÄÓ«¹â²úÂʱÈûÓнáºÏDNAµÄȾÁϸ߳ö20-30±¶£¬ËùÒÔµ±Äý½ºÖк¬ÓÐÓÎÀëµÄä廯ÒÒ¶§£¨0.5ug/ml£©Ê±£¬¿ÉÒÔ¼ì²âµ½ÉÙÖÁ10ngµÄNDAÌõ´ø¡£ä廯ÒÒ¶§¿ÉÒÔǶÈë¼î»ù·Ö×ÓÖУ¬µ¼Ö´íÅä¡£ä廯ÒÒ¶§ÊÇÇ¿ÓÕ±ä¼Á£¬¾ßÓиßÖ°©ÐÔ£¬»áÔÚ60£­70¶ÈʱÕô·¢£¨ËùÒÔ×îºÃ²»ÒªÔÚ½ºÌ«ÈȵÄʱºò¼Ó£¬»òÕßÓ¦¸Ã¼Óµ½ÒºÌåÀ0.5ug/ml£¬È¾É«°ëСʱ£©£¨µ±EB¼ÓµÃ¹ý¶àʱ£¬Ò²¿ÉÒÔÔÚÊÒÎÂÓÃË®½«ÒÑȾɫµÄÄý½º½þÅÝ20min£¬ÒÔ½µµÍδ½áºÏµÄEBÒýÆðµÄ±³¾°Ó«¹â£©¡£

ä廯ÒÒ¶§ÈÜÒºµÄ¾»»¯´¦Àí£ºÓÉÓÚä廯ÒÒ¶§¾ßÓÐÒ»¶¨µÄ¶¾ÐÔ£¬ÊµÑé½áÊøºó£¬Ó¦¶Ôº¬EBµÄÈÜÒº½øÐо»»¯´¦ÀíÔÙÐÐÆúÖã¬ÒÔ±ÜÃâÎÛȾ»·¾³ºÍΣº¦ÈËÌ彡¿µ¡£

£¨1£©¶ÔÓÚEBº¬Á¿´óÓÚ0.5mg/mlµÄÈÜÒº£¬¿ÉÈçÏ´¦Àí£º

¢Ù½«EBÈÜÒºÓÃˮϡÊÍÖÁŨ¶ÈµÍÓÚ0.5mg/ml£»

¢Ú¼ÓÈëÒ»±¶Ìå»ýµÄ0.5mol/L¸ßÃÌËá¼Ø£¬»ìÔÈ£¬ÔÙ¼ÓÈëµÈÁ¿µÄ25mol/L HCl£¬»ìÔÈ£¬ÖÃÊÒÎÂÊýСʱ£»

¢Û¼ÓÈëÒ»±¶Ìå»ýµÄ2.5mol/L NaOH£¬»ìÔȲ¢·ÏÆú¡£

£¨2£©EBº¬Á¿Ð¡ÓÚ0.5mg/mlµÄÈÜÒº¿ÉÈçÏ´¦Àí£º

¢Ù°´1mg/mlµÄÁ¿¼ÓÈë»îÐÔÌ¿£¬²»Ê±ÇáÒ¡»ìÔÈ£¬ÊÒηÅÖÃ1Сʱ£»

¢ÚÓÃÂËÖ½¹ýÂ˲¢½«»îÐÔ̼ÓëÂËÖ½ÃÜ·âºó¶ªÆú¡£

3¡¢DEPC£¨diethyl procarbonate£¬¶þÒÒ»ù½¹Ì¼Ëáõ¥£©

DEPC¿ÉÃð»î¸÷ÖÖµ°°×ÖÊ£¬ÊÇRNAøµÄÇ¿ÒÖÖÆ¼Á¡£DEPCÊÇÒ»ÖÖDZÔÚµÄÖ°©ÎïÖÊ£¬ÔÚ²Ù×÷ÖÐÓ¦¾¡Á¿ÔÚͨ·çµÄÌõ¼þϽøÐУ¬²¢±ÜÃâ½Ó´¥Æ¤·ô¡£DEPC¶¾ÐÔ²¢²»ÊǺÜÇ¿£¬µ«ÎüÈëµÄ¶¾ÐÔÊÇ×îÇ¿µÄ£¬Ê¹ÓÃʱ´÷¿ÚÕÖ¡£²»Ð¡ÐÄÕ¼µ½ÊÖÉÏ×¢ÒâÁ¢¼´³åÏ´£¬RNase AwayTMÊÔ¼Á¿ÉÒÔÌæ´úDEPC£¬²Ù×÷¼òµ¥£¬¼Û¸ñµÍ£¬ÇÒÎÞ¶¾ÐÔ¡£Ö»Ð轫RNase AwayTMÖ±½Óµ¹ÔÚ²£Á§Æ÷ÃóºÍËÜÁÏÆ÷ÃóµÄ±íÃæ£¬½þÅݺóÓÃË®³åÏ´È¥³ý£¬¼´¿ÉÒÔ¿ìËÙÈ¥³ýÆ÷Ãó±íÃæµÄRNase£¬²¢ÇÒ²»»á²ÐÁô¶ø¸ÉÈźó¼ÌʵÑé¡£

4¡¢±ûÏ©õ£°·

ÊôÖеȶ¾ÐÔÎïÖÊ£¬¿Éͨ¹ýƤ·ôÎüÊÕ¼°ºôÎüµÀ½øÈëÈËÌ壬Òò´Ë£¬ÔÚ°áÔ˺ÍʹÓÃÖбØÐë´©´÷ºÃ·À»¤Óþߣ¬Èç·À¶¾·þ£¬·À¶¾¿ÚÕÖ¼°·À¶¾ÊÖÌ׵ȡ£±ûÏ©õ£°·µÄΣº¦Ö÷ÒªÊÇÒýÆðÉñ¾­¶¾ÐÔ£¬Í¬Ê±»¹ÓÐÉúÖ³¡¢·¢Óý¶¾ÐÔ¡£Éñ¾­¶¾ÐÔ×÷ÓñíÏÖΪÖÜΧÉñ¾­ÍËÐÐÐԱ仯ºÍÄÔÖÐÉæ¼°Ñ§Ï°¡¢¼ÇÒäºÍÆäËûÈÏÖª¹¦Äܲ¿Î»µÄÍËÐÐÐԱ仯£¬ÊÔÑ黹ÏÔʾ±ûÏ©õ£°·ÊÇÒ»ÖÖ¿ÉÄÜÖ°©Îְҵ½Ó´¥ÈËȺµÄÁ÷Ðв¡Ñ§¹Û²ì±íÃ÷£¬³¤ÆÚµÍ¼ÁÁ¿½Ó´¥±ûÏ©õ£°·»á³öÏÖÊÈ˯¡¢ÇéÐ÷ºÍ¼ÇÒä¸Ä±ä¡¢»Ã¾õºÍÕð²üµÈÖ¢×´£¬°éËæÄ©ÉÒÉñ¾­²¡ÈçÊÖÌ×Ñù¸Ð¾õ¡¢³öº¹ºÍ¼¡ÈâÎÞÁ¦¡£ÀÛ»ý¶¾ÐÔ£¬²»ÈÝÒ×ÅŶ¾¡£

¾ß±¸ÒÔÏÂÈκÎÒ»ÏîÕߣ¬¿ÉÁÐΪÂýÐÔ±ûÏ©õ£°·Öж¾¹Û²ì¶ÔÏó£º

a.½Ó´¥±ûÏ©õ£°·µÄ¾Ö²¿Æ¤·ô³öÏֶູ¡¢ÊªÀä¡¢ÍÑÆ¤¡¢ºì°ß£»

b.³öÏÖÖ«¶ËÂéľ¡¢´ÌÍ´¡¢ÏÂÖ«·¦Á¦¡¢ÊÈ˯µÈÖ¢×´£»

c.Éñ¾­-¼¡µçͼÏÔʾÓпÉÒÉÉñ¾­Ô´ÐÔË𺦡£

ÖÎÁÆÔ­Ôò£º

¿ÉÓÃB×åάÉúËØ¡¢ÄÜÁ¿ºÏ¼Á£¬²¢¸¨ÒÔÌåÁÆ¡¢ÀíÁƼ°¶ÔÖ¢ÖÎÁÆ¡£ÖضÈÖж¾ÕßӦͬʱ¼Óǿ֧³ÖÁÆ·¨¡£

5¡¢N,N"-ÑǼ×Ë«±ûÏ©õ£°·

Óж¾£¬Ó°ÏìÖÐÊàÉñ¾­ÏµÍ³£¬ÇÐÎðÎüÈë·ÛÄ©¡£

6¡¢DTT£¨ Dithiothreitol£¬¶þÁòËÕÌÇ´¼£©

ºÜÇ¿µÄ»¹Ô­¼Á£¬É¢·¢ÄÑÎŵįøÎ¶¡£¿ÉÒòÎüÈë¡¢ÑÊÏ»òƤ·ôÎüÊÕ¶øÎ£º¦½¡¿µ¡£µ±Ê¹ÓùÌÌå»ò¸ßŨ¶È´¢´æÒºÊ±£¬´÷ÊÖÌ׺ͻ¤Ä¿¾µ£¬ÔÚͨ·ç³÷ÖвÙ×÷¡£

7¡¢TEMED£¨N,N,N",N"-Tetramethylethylenediamine£¬N,N,N",N"-Ëļ׻ùÒÒ¶þ°·£©

Ç¿Éñ¾­¶¾ÐÔ£¬·ÀÖ¹ÎóÎü£¬²Ù×÷ʱ¿ìËÙ£¬´æ·ÅʱÃÜ·â¡£

8¡¢PMSF£º£¨Phenylmethanesulfonyl fluoride£¬±½¼×»ù»Çõ£·ú£©

ÊÇÒ»ÖÖ¸ßÇ¿¶È¶¾ÐԵĵ¨¼îõ¥Ã¸ÒÖÖÆ¼Á¡£Ëü¶ÔºôÎüµÀð¤Ä¤¡¢ÑÛ¾¦ºÍƤ·ôÓзdz£´óµÄÆÆ»µÐÔ¡£¿ÉÒòÎüÈë¡¢ÑÊÏ»òƤ·ôÎüÊÕ¶øÖÂÃü¡£´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«ÑÛ¾µ£¬Ê¼ÖÕÔÚ»¯Ñ§Í¨·ç³÷ÀïʹÓá£ÔÚ½Ó´¥µ½µÄÇé¿öÏ£¬ÒªÁ¢¼´ÓôóÁ¿µÄË®³åÏ´ÑÛ¾¦»òƤ·ô£¬ÒÑÎÛȾµÄ¹¤×÷·þ¶ªÆúµô¡£

9¡¢Âȷ£¨CHCl3£©

¶ÔƤ·ô¡¢ÑÛ¾¦¡¢ð¤Ä¤ºÍºôÎüµÀÓд̼¤×÷Óá£ËüÊÇÒ»ÖÖÖ°©¼Á£¬¿ÉË𺦸κÍÉö¡£ËüÒ²Ò×»Ó·¢£¬±ÜÃâÎüÈë»Ó·¢µÄÆøÌå¡£²Ù×÷ʱ´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«ÑÛ¾µ²¢Ê¼ÖÕÔÚ»¯Ñ§Í¨·ç³÷Àï½øÐС£

10¡¢¼×È©£¨HCOH£©

ÓкܴóµÄ¶¾ÐÔ²¢Ò×»Ó·¢£¬Ò²ÊÇÒ»ÖÖÖ°©¼Á¡£ºÜÈÝÒ×ͨ¹ýƤ·ôÎüÊÕ£¬¶ÔÑÛ¾¦¡¢ð¤Ä¤ºÍÉϺôÎüµÀÓд̼¤ºÍËðÉË×÷ÓᣱÜÃâÎüÈëÆä»Ó·¢µÄÆûÎí¡£Òª´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«ÑÛ¾µ£¬Ê¼ÖÕÔÚ»¯Ñ§Í¨·ç³÷ÄÚ½øÐвÙ×÷¡£Ô¶ÀëÈÈ¡¢»ð»¨¼°Ã÷»ð¡£

11¡¢¼ªÄ·Èø£¨Giemsa£©

ȾÁÏÑÊÏ¿ÉÖÂÃü»òÒýÆðÑÛ¾¦Ê§Ã÷£¬Í¨¹ýÎüÈëºÍƤ·ôÎüÊÕ»áÔì³ÉÉ˺¦¡£´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«»¤Ä¿¾µ£¬ÔÚ»¯Ñ§Í¨·ç³÷Àï²Ù×÷£¬²»ÒªÎüÈëÆä·ÛÄ©¡£

12¡¢µþµªÄÆ£¨NaN3£©

¶¾ÐԷdz£´ó£¬ÄÜ×è¶Ïϸ°ûÉ«ËØµç×ÓÔËËÍϵͳ¡£¶ÔÓÚº¬ÓеþµªÄƵÄÈÜÒº£¬ÒªÃ÷È·±ê¼Ç¡£¿ÉÒòÎüÈë¡¢ÑÊÏ»òƤ·ôÎüÊÕ¶øË𺦽¡¿µ¡£´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«»¤Ä¿¾µ£¬²Ù×÷ʱҪ¸ñÍâСÐÄ¡£

13¡¢Ê®¶þÍé»ùÁòËáÄÆ£¨SDS£©

Óж¾£¬ÊÇÒ»Öִ̼¤Î²¢Ôì³É¶ÔÑÛ¾¦µÄÑÏÖØËðÉ˵ÄΣÏÕ¡£¿ÉÒòÎüÈë¡¢ÑÊÏ»òƤ·ôÎüÊÕ¶øË𺦽¡¿µ¡£´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«»¤Ä¿¾µ¡£²»ÒªÎüÈëÆä·ÛÄ©¡£

14¡¢ÈýÂÈÒÒËᣨTCA£©
  
ÇÖÈë;¾¶£ºÎüÈ롢ʳÈë¡¢¾­Æ¤ÎüÊÕ¡£
½¡¿µÎ£º¦£ºÎüÈ뱾Ʒ·Û³¾¶ÔºôÎüµÀÓд̼¤×÷Ó㬿ÉÒýÆð¿ÈËÔ¡¢ÐØÍ´ºÍÖÐÊàÉñ¾­ÏµÍ³ÒÖÖÆ¡£ÑÛÖ±½Ó½Ó´¥¿ÉÔì³ÉÑÏÖØË𺦣¬ÖØÕ߿ɵ¼ÖÂʧÃ÷¡£Æ¤·ô½Ó´¥¿ÉÖ»¯Ñ§ÐÔׯÉË¡£¿Ú·þׯÉË¿ÚÇ»ºÍÏû»¯µÀ£¬³öÏÖ¾çÁÒ¸¹Í´¡¢Å»ÍºÍÐéÍÑ¡£
  
15¡¢Triton X-100£¨¾ÛÒÒ¶þ´¼ÐÁ»ù±½»ùÃÑ£©

ÊÇÒ»ÖÖ·ÇÀë×ÓÐͱíÃæ»îÐÔ¼Á£¨»ò³ÆÈ¥ÎÛ¼Á£©¡£Äܹ»ÒýÆðÑÏÖØµÄÑÛ¾¦´Ì¼¤ºÍׯÉË¡£¿ÉÒòÎüÈë¡¢ÑÊÏ»òƤ·ôÎüÊÕ¶øÊܺ¦¡£´÷ºÏÊʵÄÊÖÌ׺ͻ¤Ä¿¾µ¡£

16¡¢¹ýÁòËáï§

¶Ôð¤Ä¤ºÍÉϺôÎüµÀ×éÖ¯¡¢ÑÛ¾¦ºÍƤ·ôÓм«´óΣº¦ÐÔ¡£ÎüÈë¿ÉÖÂÃü¡£²Ù×÷ʱ´÷ºÏÊʵÄÊÖÌס¢°²È«ÑÛ¾µºÍ·À»¤·þ¡£Ê¼ÖÕÔÚͨ·ç³÷Àï²Ù×÷£¬²Ù×÷Íêºó³¹µ×Ï´ÊÖ¡£

17¡¢TRIzol

º¬Óж¾ÎïÖʱ½·Ó£¬ÈçÆ¤·ô½Ó´¥TRIzol£¬ÇëÁ¢¼´ÓôóÁ¿È¥¹¸¼ÁºÍË®³åÏ´£¬ÈçÈÔÓв»ÊÊ£¬Ç뾡¿ì¾ÍÒ½¡£Èç¹ûÖ»ÊÇÉÙÁ¿½Ó´¥£¬²¢´¦ÀíºóÖ¢×´¼õÇᣬ¹À¼ÆÎÊÌâ²»´ó¡£
»Ø¸´´ËÂ¥

» ÊÕ¼±¾ÌûµÄÌÔÌûר¼­ÍƼö

³¬¼¶µçÈÝÆ÷¼°¸÷ÖÖ¿ÆÑо«»ª Better Resouces

» ²ÂÄãϲ»¶

» ±¾Ö÷ÌâÏà¹ØÉ̼ÒÍÆ¼ö: (ÎÒÒ²ÒªÔÚÕâÀïÍÆ¹ã)

» ±¾Ö÷ÌâÏà¹Ø¼ÛÖµÌùÍÆ¼ö£¬¶ÔÄúͬÑùÓаïÖú:

ÒÑÔÄ   »Ø¸´´ËÂ¥   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû

remember921

ľ³æ (СÓÐÃûÆø)


¡ï¡ï¡ï¡ï¡ï ÎåÐǼ¶,ÓÅÐãÍÆ¼ö

ºÃ¶«Î÷°¡ ÍòÒ»Öж¾Á˾ÍÂé·³ÁË
5Â¥2010-12-01 13:12:00
ÒÑÔÄ   »Ø¸´´ËÂ¥   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû
²é¿´È«²¿ 8 ¸ö»Ø´ð

315492573

Òø³æ (³õÈëÎÄ̳)


¡ï Ò»ÐǼ¶,Ò»°ã

ºÜ¶àÇé¿öÏ ʵÑé¹ý³ÌÖв¢Ã»ÔõôעÒ⣬°¦¡­¡­
4Â¥2010-11-21 08:59:45
ÒÑÔÄ   »Ø¸´´ËÂ¥   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû

ÏþÇÙ±¦±¦

ľ³æ (ÖøÃûдÊÖ)


¡ï¡ï¡ï¡ï¡ï ÎåÐǼ¶,ÓÅÐãÍÆ¼ö

ʵÑéÊÒ»¹ÓкöàÓÃÆ·Óж¾ÄØ£¬·À²»Ê¤·À°¡£¬Ö»ÄÜŬÁ¦·À°¡
8Â¥2014-09-15 11:14:25
ÒÑÔÄ   »Ø¸´´ËÂ¥   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû
¡î ÎÞÐǼ¶ ¡ï Ò»ÐǼ¶ ¡ï¡ï¡ï ÈýÐǼ¶ ¡ï¡ï¡ï¡ï¡ï ÎåÐǼ¶
×î¾ßÈËÆøÈÈÌûÍÆ¼ö [²é¿´È«²¿] ×÷Õß »Ø/¿´ ×îºó·¢±í
[¿¼ÑÐ] 310Çóµ÷¼Á +11 666ÕæºÃ 2026-04-11 12/600 2026-04-12 06:05 by hmn_wj
[¿¼ÑÐ] 280Çóµ÷¼Á +13 wzzzÍõ 2026-04-09 13/650 2026-04-12 00:31 by ÓÂÅʸ߷å0126
[¿¼ÑÐ] 22ר˶Çóµ÷¼Á +6 haoyunÉϰ¶ 2026-04-11 8/400 2026-04-11 23:21 by labixiaoqiao
[¿¼ÑÐ] µ÷¼Á +6 ÇàµÆ²»¸º 2026-04-09 6/300 2026-04-11 20:35 by dongdian1
[¿¼ÑÐ] 343Çóµ÷¼Á +9 Íõ¹ú˧ 2026-04-10 9/450 2026-04-11 20:31 by dongdian1
[¿¼ÑÐ] 296Çóµ÷¼Á +13 Íô£¡£¿£¡ 2026-04-10 15/750 2026-04-11 10:31 by ÄæË®³Ë·ç
[¿¼ÑÐ] 293Çóµ÷¼Á +6 ÓÂÔ¶¿â°®314 2026-04-08 6/300 2026-04-11 10:08 by zhq0425
[¿¼ÑÐ] ²ÄÁÏרҵ344Çóµ÷¼Á +16 hualkop 2026-04-10 21/1050 2026-04-10 17:28 by laoshidan
[¿¼ÑÐ] 336²ÄÁÏÓ뻯¹¤085600Çóµ÷¼Á +21 Ë®ÐǼÇinfp 2026-04-05 24/1200 2026-04-10 15:28 by luoyongfeng
[¿¼ÑÐ] 347²ÄÁÏר˶Çóµ÷¼Á +19 zj8215216 2026-04-06 19/950 2026-04-10 09:36 by 690616278
[¿¼ÑÐ] ²ÄÁϹ¤³Ìµ÷¼Á +12 СÁõͬѧ߹߹ 2026-04-06 13/650 2026-04-09 17:07 by luoyongfeng
[¿¼ÑÐ] Çó»úеר˶297µÚ¶þÅúµ÷¼Á +5 ʰÆâ12¡£ 2026-04-08 5/250 2026-04-09 16:43 by Ôʵ±ÊʶÈ
[¿¼ÑÐ] 265Çóµ÷¼Á +4 ·ç˵ËýÔçÍüÁË 2026-04-07 4/200 2026-04-09 13:59 by onlyÖÜ
[¿¼ÑÐ] Ò»Ö¾Ô¸Ö£ÖÝ´óѧ085600Çóµ÷¼Á +21 ³ÔµÄ²»ÉÙ 2026-04-05 24/1200 2026-04-08 16:47 by sunhuadong
[¿¼ÑÐ] 11408 325·Ö +3 jgtxuxgkx 2026-04-07 3/150 2026-04-07 23:10 by lbsjt
[¿¼ÑÐ] 277Çóµ÷¼Á ÊýÒ»104·Ö +9 Æ¿×ÓPZ 2026-04-05 14/700 2026-04-07 17:52 by À¶ÔÆË¼Óê
[¿¼ÑÐ] ²ÄÁϵ÷¼Á +17 СÁõͬѧ߹߹ 2026-04-06 18/900 2026-04-07 11:41 by Ê«Óë×ÔÓÉ
[¿¼ÑÐ] 285Çóµ÷¼Á +5 mapmath 2026-04-06 6/300 2026-04-06 17:18 by À¶ÔÆË¼Óê
[¿¼ÑÐ] ¹¤¿Æ370Çóµ÷¼Á +3 äçÐļ弦µ° 2026-04-05 3/150 2026-04-06 10:55 by ÕâÊÇÒ»¸öÎÞÁĵÄê
[¿¼ÑÐ] 332Çóµ÷¼Á +17 ССÃÏ... 2026-04-05 18/900 2026-04-06 09:51 by À¶ÔÆË¼Óê
ÐÅÏ¢Ìáʾ
ÇëÌî´¦ÀíÒâ¼û