Znn3bq.jpeg
²é¿´: 482  |  »Ø¸´: 1
µ±Ç°Ö÷ÌâÒѾ­´æµµ¡£

zycly86

Í­³æ (СÓÐÃûÆø)

[½»Á÷] ¡¾·ÖÏí¡¿·Ö×ÓʵÑéÊÒÓж¾ÊÔ¼Á

1.DMSO£ºDMSOÊǶþ¼×»ùÑÇí¿£¬ ÓÃ;¹ã·º¡£ÓÃ×÷ÒÒȲ¡¢·¼Ìþ¡¢¶þÑõ»¯Áò¼°ÆäËûÆøÌåµÄÈܼÁÒÔ¼°ëæÂÚÏËά·ÄË¿ÈܼÁ¡£ÊǼ´ÈÜÓÚË®ÓÖÈÜÓÚÓлúÈܼÁµÄ¼«ÎªÖØÒªµÄ·ÇÖÊ×Ó¼«ÐÔÈܼÁ¡£¶ÔƤ·ôÓм«Ç¿µÄÉøÍ¸ÐÔ£¬ÓÐÖúÓÚÒ©ÎïÏòÈËÌåÉøÍ¸¡£Ò²¿É×÷ΪũҩµÄÌí¼Ó¼Á¡£Ò²ÊÇÒ»ÖÖÊ®·ÖÖØÒªµÄ»¯Ñ§ÊÔ¼Á;
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£¼ûµÄΪ¶ñÐÄ¡¢Å»Í¡¢Æ¤Õî¼°ÔÚÆ¤·ô¡¢ºÍºô³öµÄÆøÌåÖз¢³ö´óËâ¡¢Ñó´Ð¡¢ÄµòÃζ¡£
ÎüÈ룺 ¸ß»Ó·¢Å¨¶È¿ÉÄܵ¼ÖÂÍ·Í´£¬ÔÎÑ£ºÍÕò¾²¡£
Ƥ·ô£º Äܹ»×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¡£
ÎüÊÕ£º ÎüÊÕΣÏÕÐԺܵ͡£

2.EB£ºEB£¨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»¿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໥×÷Ó᣸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´¡£ÓÉÓÚä廯ÒÒ¶§-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¶ÈÊÇÕô·¢£¬£¨ËùÒÔ×îºÃ²»ÒªÔÚ½ºÌ«ÈȵÄʱºò¼Ó£¬»òÕßÓ¦¸Ã¼Óµ½ÒºÌåÀ0.5ug/ml£¬È¾É«°ëСʱ£¬£©£¨µ±EB¼ÓµÃ¹ý¶àʱ£¬Ò²¿ÉÒÔÔÚÊÒÎÂÓÃË®½«ÒÑȾɫµÄÄý½º½þÅÝ20minÒÔ½µµÍδ½áºÏµÄEBÒýÆðµÄ±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 KMnO4£¬»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£»
¢Ú ÓÃÂËÖ½¹ýÂ˲¢½«»îÐÔ̼ÓëÂËÖ½ÃÜ·âºó¶ªÆú¡£

¼û¹â·Ö½â£¿£¿Ëû˵Ëû¾­³£ÂãÊÖ×¥½ºµÄ£¬Ö»ÓÐѧÉúÔÚµÄʱºò²Åʾ·¶ÐԵĴ÷Ò»ÏÂÊÖÌס£µ±Ê±ÎÒ¿´Ê±¶¼³ÔÁËÒ»¾ª£¬µ«ÌýËûÒ»½âÊÍ£¬Ò²¾õµÃÓеãµÀÀíµÄ¡£ Ëû˵£¬EBÈç¹û½Ó´¥µ½ÄãµÄƤ·ô£¬Ò»°ã¶¼ÊÇÊÖÁË£¬ÆäÉ˺¦µÄ¶¼ÊÇ±íÆ¤Ï¸°û£¬¶øÕâÖ»ÊÇÒ»²ãËÀÍöÁ˵Äϸ°û£¬ÆäÉ˺¦ºÜÄÑ´ïµ½ÕæÆ¤²ã¡£
ÏÖÔÚÒѾ­ÓкܶàÎÞ¶¾µÄȼÁÏÂôÁË£¬ÀýÈçGODview

3.DEPC

DEPC¼´¶þÒÒ»ù½¹Ì¼Ëáõ¥£¨diethylprocarbonate£©£¬¿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£°·£ºÊôÖеȶ¾ÐÔÎïÖÊ¡£¿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±¸ÒÔÏÂÈκÎÕߣ¬¿ÉÁÐΪÂýÐÔ±ûÏ©õ£°·Öж¾¹Û²ì¶ÔÏó:
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
¿ÉÓÃB×åάÉúËØ¡¢ÄÜÁ¿ºÏ¼Á£¬²¢¸¨ÒÔÌåÁÆ¡¢ÀíÁƼ°¶ÔÖ¢ÖÎÁÆ¡£ÖضÈÖж¾ÕßӦͬʱ¼Óǿ֧³ÖÁÆ·¨¡£

5. NN-ÑǼ×Ë«±ûÏ©õ£°·£¬Óж¾£¬Ó°ÏìÖÐÊàÉñ¾­ÏµÍ³£¬ÇÐÎðÎüÈë·ÛÄ©¡£

6 DTT ¶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 TEMEDÇ¿Éñ¾­¶¾ÐÔ,·ÀÖ¹ÎóÎü,²Ù×÷ʱ¿ìËÙ,´æ·ÅʱÃÜ·â.
8¡£ PMSF£º±½¼×»ù»Çõ£·ú[(PMSF)£¬C7H7FO2S»òC6H5CH2SO2F]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. ÂÈ·Â(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·ôÎüÊÕÊÇÓж¾µÄ¡£Æä¿ÉÄܵÄΣÏÕÊDz»¿ÉÄæµÄЧӦ¡£´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«»¤Ä¿¾µ¡£ÔÚ»¯Ñ§Í¨·ç³÷Àï²Ù×÷£¬²»ÒªÎüÈëÆä·ÛÄ©¡£

12. µþµªÄÆ(NaN3)¶¾ÐԷdz£´ó¡£Ëü×è¶Ïϸ°ûÉ«ËØµç×ÓÔËËÍϵͳ¡£º¬ÓеþµªÄƵÄÈÜÒºÒª±ê¼ÇÇå³þ¡£¿ÉÒòÎüÈë¡¢ÑÊÏ»òƤ·ôÎüÊÕ¶øË𺦽¡¿µ¡£´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«»¤Ä¿¾µ£¬²Ù×÷ʱҪ¸ñÍâСÐÄ¡£

13Ê®¶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. ÈýÂÈÒÒËá(TCA)ÓкÜÇ¿µÄ¸¯Ê´ÐÔ¡£´÷ºÏÊʵÄÊÖÌ׺Ͱ²È«·ÀÄ¿¾µ¡£

15. Triton X-100ÒýÆðÑÏÖØµÄÑÛ¾¦´Ì¼¤ºÍׯÉË¡£¿ÉÒòÎüÈë¡¢ÑÊÏ»òƤ·ôÎüÊÕ¶øÊܺ¦¡£´÷ºÏÊʵÄÊÖÌ׺ͻ¤Ä¿¾µ¡£

16. ¹ýÁòËáï§[(NH4)2S2O8]¶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. Trizolº¬Óж¾ÎïÖʱ½·Ó,ÈçÆ¤·ô½Ó´¥Trizol,ÇëÁ¢¼´ÓôóÁ¿È¥¹¸¼ÁºÍË®³åÏ´,ÈçÈÔÓв»ÊÊ,ÇëÌýȡҽÉúÒâ¼û¡£Èç¹ûÖ»ÊÇÉÙÁ¿½Ó´¥£¬²¢´¦ÀíºóÖ¢×´¼õÇᣬ¹À¼ÆÎÊÌâ¾Í²»´óÁË¡£

×ÏÍâ¹â»ò×ÏÍâÏß¿ÉËðÉËÑÛÊÓÍøÄ¤¡£ÇÐÎðÓÃÂãÑÛºÍûÓзÀ»¤×°ÖõÄ×ÏÍâ¹âÔ´¡£ÔÚʵÑéÊÒÀï³£ÓõÄ×ÏÍâ¹âÔ´°üÀ¨ÊÖÌáʽ×ÏÍâµÆºÍ×ÏÍâ͸ÉäÒÇ¡£Ö»ÄÜͨ¹ýÎüÊÕÓк¦²¨³¤µÄÂËÆ¬»ò°²È«²£Á§Æ¬²ÅÄܹ۲졣×ÏÍâÏßÒ²ÊÇÓÕ±ä¼ÁºÍÖ°©µÄ¡£ÎªÊ¹±©Â¶¼õÉÙµ½×îµÍÏÞ¶È£¬È·±£×ÏÍâ¹âÔ´Òª²ÉÓÃÊʵ±·À»¤×°Öá£ÔÚ×ÏÍâ¹âϲÙ×÷ʱҪ´÷ºÏÊʵÄÔ¤·ÀÐÔÊÖÌס£

»¹Òª×¢Òâ¾Æ¾«µÆµÄʹÓã¡Ð¡ÐÄ×Å»ð£¡
»Ø¸´´ËÂ¥

» ÊÕ¼±¾ÌûµÄÌÔÌûר¼­ÍƼö

¿ÆÑÐ×ÊÁÏ

» ²ÂÄãϲ»¶

» ±¾Ö÷ÌâÏà¹ØÉ̼ÒÍÆ¼ö: (ÎÒÒ²ÒªÔÚÕâÀïÍÆ¹ã)

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

1

лл·ÖÏí£¡
2Â¥2010-02-20 17:33:48
ÒÑÔÄ   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû
Ïà¹Ø°æ¿éÌø×ª ÎÒÒª¶©ÔÄÂ¥Ö÷ zycly86 µÄÖ÷Ìâ¸üÐÂ
×î¾ßÈËÆøÈÈÌûÍÆ¼ö [²é¿´È«²¿] ×÷Õß »Ø/¿´ ×îºó·¢±í
[¿¼ÑÐ] µç×ÓÐÅÏ¢279Çóµ÷¼Á£¬ÓÐÊé¶Á¾ÍÐÐ +8 wwwooden 2026-04-08 11/550 2026-04-11 20:22 by cq2548
[¿¼ÑÐ] 331Çóµ÷¼Á +4 Íõ¹ú˧ 2026-04-11 4/200 2026-04-11 20:19 by À¶ÔÆË¼Óê
[˶²©¼ÒÔ°] ÐÂÒ»´úµç×ÓÐÅÏ¢294Çóµ÷¼Á ²»ÌôѧУ +6 Ytyt11 2026-04-09 7/350 2026-04-11 10:52 by AAССľ³æ
[¿¼ÑÐ] 296Çóµ÷¼Á +13 Íô£¡£¿£¡ 2026-04-10 15/750 2026-04-11 10:31 by ÄæË®³Ë·ç
[¿¼ÑÐ] 22408 327·ÖÇóµ÷¼Á +4 ÔÏ·çkon 2026-04-10 4/200 2026-04-11 09:51 by Öí»á·É
[¿¼ÑÐ] Ò»Ö¾Ô¸¾©Çø985£¬085401£¬Óë±¾¿Æ×¨ÒµÒ»Ö£¬µç×ÓÐÅÏ¢¹¤³Ì£¬ +4 Ñô¹â¿ªÀʵÄÄк¢ 2026-04-10 4/200 2026-04-10 18:27 by shenrf
[¿¼ÑÐ] 307Çóµ÷¼Á +8 tzq94092 2026-04-10 8/400 2026-04-10 17:33 by 286640313
[¿¼ÑÐ] ±¾9 Ò»Ö¾Ô¸Î÷¹¤´ó085601 324Çóµ÷¼Á +5 wysyjs25 2026-04-10 5/250 2026-04-10 16:57 by luoyongfeng
[ÂÛÎÄͶ¸å] mdpiСÐÞrvrʱ¼äËÄÎåÌìÁË 20+3 ¹þ¹þhigh 2026-04-08 5/250 2026-04-10 16:02 by ±±¾©À³ÒðÈóÉ«
[¿¼ÑÐ] ¿¼Ñе÷¼Á-²ÄÁÏÀà-284 +28 Ïë»»ÊÖ»ú²»Ïë½âÊ 2026-04-08 28/1400 2026-04-09 20:08 by µ¹Êý321?
[¿¼ÑÐ] 337Çóµ÷¼Á +4 Gky09300550£¬ 2026-04-09 4/200 2026-04-09 17:18 by ÅÁ¶ûÂíÀ­ÌØ
[¿¼ÑÐ] ²ÄÁϹ¤³Ì322 +18 ¹þ¹þ¹þºðºðºð¹þ 2026-04-07 19/950 2026-04-09 10:44 by cymywx
[¿¼ÑÐ] Ò»Ö¾Ô¸985³õÊÔ354·ÖÉúÎïµ÷¼Á +3 031001 2026-04-06 3/150 2026-04-09 00:30 by Evan_Liu
[¿¼ÑÐ] 353Çóµ÷¼Á +8 Çç¿ÕÍòÀïair 2026-04-07 8/400 2026-04-09 00:18 by GouQ
[¿¼ÑÐ] ÉúÎïÒ½Ò©µ÷¼Á£üSCIÖпÆÔºÈýÇøÒ»×÷+¶àÏî¿ÆÑгɹû +8 likangxing 2026-04-07 11/550 2026-04-08 00:02 by lys0704
[¿¼ÑÐ] 323Çóµ÷¼Á +3 ÁÖzlu 2026-04-07 4/200 2026-04-07 23:21 by lbsjt
[¿¼²©] ²©Ê¿ÉêÇë +3 IQwQl 2026-04-05 3/150 2026-04-07 20:31 by greychen00
[¿¼ÑÐ] ÉúÎïѧѧ˶Çóµ÷¼Á£º351·ÖÒ»Ö¾Ô¸ÄϾ©Ê¦·¶´óѧÉúÎïѧרҵ +6 ¡­¡«¡¢Íõ¡­¡« 2026-04-06 7/350 2026-04-06 18:54 by macy2011
[¿¼ÑÐ] 285Çóµ÷¼Á +5 mapmath 2026-04-06 6/300 2026-04-06 17:18 by À¶ÔÆË¼Óê
[¿¼ÑÐ] ¹¤¿Æ277·ÖÇóµ÷¼Á²ÄÁÏ +8 ÉÏÁËÉÏÁËÉÏŶ 2026-04-05 9/450 2026-04-05 13:05 by wwytracy
ÐÅÏ¢Ìáʾ
ÇëÌî´¦ÀíÒâ¼û