• ÇíÖ¬ÌÇÄý½ºµçÓ¾Ô­Àí/ʵÑé²½Öè/ÌØµã/Ïà¹ØFAQ

    ±¾ÎÄÖ÷Òª½éÉÜÁËÇíÖ¬ÌÇÄý½ºµçÓ¾µÄÔ­Àí¼°ÊµÑé²Ù×÷²½Ö裬¼°µçÓ¾ºóDNAµÄ»ØÊÕ·½·¨£»ÇíÖ¬ÌÇÄý½ºµÄÌØµã¼°µçӾʵÑéÏà¹ØÊÔ¼ÁµÄÅäÖü°ÇíÖ¬ÌÇÄý½ºµçÓ¾³£¼ûÎÊÌâ·ÖÎö¡£

    ÇíÖ¬ÌÇÄý½ºµçÓ¾ÑùÆ·ÖÆ±¸

    ´ó³¦¸Ë¾úÖÊÁ£DNAÄ£°åÌáÈ¡£º

    1¡¢½Ó1%º¬ÖÊÁ£µÄ´ó³¦¸Ë¾úϸ°ûÓÚ2ml?LBÅàÑø»ù

    2¡¢37¡æÕñµ´ÅàÑø¹ýÒ¹

    3¡¢È¡100ul¾úÌåÓÚ2mlEp¹ÜÖУ¬4000rpmÀëÐÄ3min£¬ÆúÉÏÇå

    4¡¢¼Ó100ulÈÜÒºA(1£¥ÆÏÌÑÌÇ50mM/L?EDTA?pH8.0£¬25mM/L?Tris-HCl?pH8.0)³ä·Ö»ìºÏ

    5¡¢¼ÓÈë200ulÈÜÒº?B(0.2?mM/L?NaOH£¬1£¥?SDS)£¬ÇáÇᷭת»ìÔÈ£¬ÖÃÓÚ±ùÔ¡5?min

    6¡¢¼ÓÈë150ulÔ¤ÀäÈÜÒºC(5?mol/L?KAc£¬pH4.8)£¬ÇáÇᷭת»ìÔÈ£¬ÖÃÓÚ±ùÔ¡5?min

    7¡¢10000rpmÀëÐÄ20min£¬È¡ÉÏÇå

    8¡¢¼ÓÈëµÈÌå»ýµÄÒìÎì´¼£¬»ìÔȺóÓÚ60¡æ¾²ÖÃ10min

    9¡¢ÔÙ´Î10000rpmÀëÐÄ20min£¬ÆúÉÏÇå

    10¡¢ÓÃ500ul70£¥µÄÒÒ´¼Ï´µÓ£¬³é¸ÉËùÓÐÒºÌå

    11¡¢´ý³Áµí¸ÉÔÈÜÓÚ5ulµÄTE»º³åÒºÖУ¨Í¨³£¿É»ñµÃ3-5ugµÄDNA£©

    ÕæºËϸ°ûDNAÄ£°åÌáÈ¡£º

    ÕæºËϸ°ûDNAµÄÌáÈ¡ÊǽøÐй¦ÄܺͽṹÑо¿µÄÖØÒªÒ»²½£¬ÕæºËDNAÌáȡҪ±£Ö¤DNAµÄÍêÕûÐÔ£¨²»¶ÏÁÑ£©¡£ÕæºËDNA¿ÉÒÔ´ÓÅàÑøµÄϸ°ûÐÂÏʵÄ×éÖ¯ÖÐÌáÈ¡¡£Í¨³£¶¼ÊDzÉÓõ°°×øÏû»¯ºóÓ÷ӳéÌá»ñµÃ¡£²»Í¬À´Ô´µÄ²ÄÁϽøÐв»Í¬µÄÔ¤´¦Àí£¬µ«ÊǺóÐøÌáÈ¡DNAµÄ·½·¨ÊÇÀàËÆµÄ£¬ÌáÈ¡µÄÔ­Ôò¾ÍÊÇ£¬¾¡Á¿¼õÉÙ¶ÔDNAµÄÆÆ»µ»ò½µ½â£¬±£³ÖÆäÍêÕûÐÔ¡£¾ßÌåÌáÈ¡²½ÖèÈçÏ£º

    £¨Ò»£©²ÄÁÏÔ¤´¦Àí£º

    ÅàÑøÏ¸°ûÓûº³åҺϴµÓºóÀëÐÄ£¨4000rpm/5min£©È¥³ýÉÏÇ壬µÃµ½Ï¸°û³ÁµíÎï¼ÓÈë10±¶Ìå»ýµÄÁѽ⻺³åÒº£¬55¡æË®Ô¡1-2Сʱ£»

    ×éÖ¯£¨×éÖ¯Òª¼ôËéÔȽ¬£©¼ÓÈë1.5mlµ½ÀëÐĹÜÖУ¬¼ÓÈë20%µÄSDS?25ul£¬µ°°×øK£¨2mg/ml£©25ul£¬È»ºó»ìÔÈ£¬60¡æË®Ô¡1-3Сʱ£»

    £¨¶þ£©DNAµÄÌáÈ¡£¨ÊÊÓÃÓÚ´¦ÀíºóµÄϸ°û»ò×éÖ¯£©

    1. Ô¤´¦ÀíµÄÑùÆ·¼ÓÈë±¥ºÍ·Ó£¬ÇáÇá»ìÔȼ¸·ÖÖÓºóÀëÐÄ£¨5000rpm/10min£©£¬È¥ÉϲãË®Ïࣻ
    2. ¼ÓÈëµÈÌå»ýµÄ±¥ºÍ·Ó£¬»ìÔÈ£¬ÔÙ´ÎÀëÐÄ£¬È¥ÉϲãË®ÏֱࣻÖÁÈ¥ÉϲãË®Ïà±äµÃ³ÎÇ壬²»È»¿ÉÖØ¸´´Ë²Ù×÷£»
    3. ¼ÓÈëÊ®·ÖÖ®Ò»µÄ3M´íËáÄÆ£¨pH2.5£©HE?2.5±¶Ìå»ýµÄÎÞË®ÒÒ´¼£¬ÇáÇá»ìÔÈ£»
    4. ³öÏÖÐõ×´ÎïºóÀëÐÄ£¨5000rpm/10min£©£¬ÆúÉÏÇ壻ÓÃ75%ÒÒ´¼Ï´µÓ³Áµí£¬ÔÙ´ÎÀëÐÄ£¬ÆúÉÏÇ壻
    5. ÊÒÎÂÏÂÁÀ¸É£¨´ýÒÒ´¼»Ó·¢£©£¬¼ÓÈë100ul?TEÈܽ⼴¿É¡£

    ÌáÈ¡DNAµÄÎïÆ·ÐèÒª¸ßѹÃð¾ú£¬ÒÔ·ÀÖ¹´øÈëÆäËûºËËáÎÛȾ£»ÊÔ¼ÁͬÑù¸ßѹÃð¾ú£»

    PCRÀ©Ôö

    PCRÀ©ÔöʵÑé²½ÖèÏê¼û£ºPCR±ê×¼²Ù×÷Á÷³Ì¡£

    ÇíÖ¬ÌÇÄý½ºµçÓ¾Ô­Àí

    ÇíÖ¬ÌÇÄý½ºµçÓ¾£¬ÊÇÒÔÇíÖ¬ÌÇΪ½éÖÊ£¬¶Ô²»Í¬´óСµÄDNA»òRNAʵÏÖ·ÖÀëµÄÒ»ÖÖµçÓ¾·½·¨¡£ÇíÖ¬ÌÇÊÇÒ»ÖÖ¶àÌÇ£¬¾ßÓÐÇ×Ë®ÐÔ£¬µ«ÊDz»´øµçºÉ¡£Ê¹µÃDNAÔÚ¼îÐÔÌõ¼þÏÂʹÆä´ø¸ºµçºÉ£¨pH8.0µÄ»º³åÒº£©£¬ÔÚµçÁ÷×÷ÓÃÏ£¬ÒÔÇíÖ¬ÌÇÄý½ºÎª½éÖÊ£¬Óɸº¼«ÏòÕý¼«Òƶ¯£¬¸ù¾Ý²»Í¬µÄDNA·Ö×ÓÆ¬¶ÎµÄ´óСºÍÐÎ×´²»Í¬£¬Ôڵ糡ÖÐÓ¾¶¯µÄËÙÂÊÒ²²»Ïàͬ£¬Í¬Ê±ÔÚÑùÆ·ÖмÓÈëȾÁÏ£¨ÈçEB»ò»¨ÇàËØÀàȾÁÏ£©Äܹ»ºÍDNA·Ö×Ó¼äÐγÉÂçºÏÎ¾­¹ý×ÏÍâÕÕÉ䣬¿ÉÒÔ¿´µ½DNAµÄλÖ㨱ȶÔmarker¿ÉÖª·Ö×ÓÁ¿´óС£©£¬´Ó¶ø´ïµ½·ÖÀë¡¢¼ø¶¨µÄÄ¿µÄ¡£Èçͼ1ΪÇíÖ¬ÌÇÄý½ºµçÓ¾Ô­Àí¡£

    ÇíÖ¬ÌÇÄý½ºµçÓ¾Ô­Àí

    ͼ1£ºÇíÖ¬ÌÇÄý½ºµçÓ¾Ô­Àí

    ÇíÖ¬ÌÇÄý½ºµçÓ¾Óŵã

    1. ÇíÖ¬ÌÇÄý½ºÊǾßÓдóÁ¿Î¢Ð¡¿×µÀµÄ»ùÖÊ£¬Æä¿×¾¶³ß´çÈ¡¾öÓÚËüµÄŨ¶È¡£0.075ÇíÖ¬ÌǵĿ׾¶Îª800nm,0.16µÄ¿×¾¶Îª500nm£¬1%µÄÇíÖ¬Ìǿ׾¶Îª150nm£¬Õâ¶¼Êdz£ÓõÄÇíÖ¬ÌÇÄý½ºµÄŨ¶È£¬¿ÉÒÔ·ÖÀë0.1-60kb×óÓÒµÄDNA£¬´ó¿×ÐÔÓÐÀûÓÚÃâÒ߹̶¨¡¢ÃâÒßµçÓ¾ºÍ΢Á¿ÖƱ¸£»
    2. ÇíÖ¬ÌǾßÓнϸ߻úеǿ¶È£¬ÔÊÐíÔÚ1%»ò¸üµÍµÄŨ¶ÈÏÂʹÓã»ÇÒÓë±ðµÄÉúÎïÖ»´æÔÚ΢ÈõµÄ½áºÏ£»
    3. ÇíÖ¬ÌÇÎÞ¶¾£¬ÇíÖ¬ÌǽºÄý¹Ì¹ý³Ì²»»á·¢Éú×ÔÓÉ»ù¾ÛºÏ£¬ÎÞÐè´ß»¯¼Á£»
    4. ÇíÖ¬Ìǽº¾ßÓÐÈÈ¿ÉÄæÐÔ£¬µÍÈÛµãÒ×ÓÚ»ØÊÕÑùÆ·£¬¿ÉÓÃÓÚ¶ÔζÈÃô¸Ð²ÄÁϵÄÖÆ±¸£»
    5. Ò×ÓÚ±£´æ£¬ÊǸßÁéÃô·ÅÉä×ÔÏÔÓ°µÄÀíÏë²ÄÁÏ¡£

    ÇíÖ¬ÌÇÄý½ºµçÓ¾²½Öè

    ÇíÖ¬ÌÇÄý½ºµçÓ¾ÒÇÆ÷

    ͼ2£ºÇíÖ¬ÌÇÄý½ºµçÓ¾ÒÇÆ÷

    1. ÓÃÕôÁóË®½«Öƽº¹¤¾ßÏ´¸É¾»£¬×¼±¸ºÃÖÆ½ºÆ½°å£¬ÓÃÇíÖ¬½«Ä£¾ß±ßÔµ·â±Õ£¬¼ÜºÃÊá×Ó£»
    2. ¸ù¾ÝÒª·ÖÀëµÄÑùÆ·£¨DNA£©´óСÅäÖÆºÏÊÊŨ¶ÈµÄÇíÖ¬ÌÇÄý½º¡£×¼È·µÄ³ÆÈ¡Ò»¶¨Á¿µÄÇíÖ¬ÌǸɷۣ¬¼ÓÈëµ½ºÏÊÊÌå»ýµÄ×¶ÐÎÆ¿ÖУ¬¼ÓÈëÒ»¶¨Á¿µÄµçÓ¾»º³åÒº£¨30ml×óÓÒTAE£©£»·ÅÈ뵽΢²¨Â¯ÄÚ¼ÓÈÈÈÚ»¯£¬ÀäȴƬ¿Ì£¨²»ÌÌÊÖ¼´¿É£©£»
    3. ÈÚ»¯ºóµÄÇíÖ¬ÈÜÒºÒ¡»Î»ìÔÈ£¬µ¹ÈëµçÓ¾²ÛÖУ¬µÈÆäÄý¹Ì£»
    4. ÊÒÎÂÏÂÄý¹Ì30-40min×óÓÒ£¬Ð¡ÐĵİγöÊá×Ó£¬È¡³öÄý¹ÌºÃµÄÄý½º·ÅÈëµçÓ¾²ÛÄÚ£¬×¼±¸ÉÏÑù£»
    5. ÔÚµçÓ¾²ÛÖе¹ÈëµçÓ¾»º³åÒº£¨TAE£©£»Ã»¹ý½ºÃæ1mm×óÓÒ£¬È¥³ý½º¿×Ä򵀮øÅÝ£»
    6. ÉÏÑù£¬5ulDNAÑùÆ·¼ÓÈë1ulÉÏÑù»º³åÒº£¨loading?buffer£©ºÍ1ulµÄȾÁÏ£¬ÓÃÇÀ»ìÔȺó¼ÓÈëµ½Äý½º¿×ÄÚ£»
    7. ÉÏÑù½áÊø£¬½ÓͨµçÔ´£¬ºìÉ«Õý¼«£¬ºÚÉ«¸º¼«£¬DNAÑùÆ·Óиº¼«ÍùÕý¼«Ô˶¯£¬¼ÓÑù¿×²àΪ¸º£¬£¬40-50vµçѹ£¬µçѹ30Ãô×óÓÒ¼´¿É£¨<40min£©£»
    8. µçѹ½áÊø£¬¹Ø±ÕµçÔ´£¬Äý½º³ÉÏñÒÇÉϹ۲ìµçѹλÖò¢±È¶ÔmarkerÅжϴóС¡£

    ÇíÖ¬ÌÇÄý½ºÅä֯ō¶ÈÓëDNAµÄ×î¼Ñ·ÖÀ뷶Χ

    ÇíÖ¬ÌÇŨ¶È/% 0.3 0.6 0.7 0.9 1.2 1.5 2.0
    DNA´óС/kb 5-60 1-20 0.8-10 0.5-7 0.4-6 0.2-4 0.1-3

    ÇíÖ¬ÌÇÄý½ºµçÓ¾Ïà¹ØÊÔ¼ÁÅäÖÃ

    £¨Ò»£©50?x?TAE?buffer£¨pH8.5£©
    ÔÚ1LÉÕ±­ÖÐ׼ȷ³ÆÈ¡Tris?242g£¬EDTA£¨Na£©37.2g£¬¼ÓÈë800mlµÄÈ¥Àë×ÓË®£¬³ä·Ö½Á°èÈܽ⣬¼ÓÈë57.1mlµÄÒÒËᣬ³ä·Ö½Á°è£¬µ÷pH8.5ºó£¬¶¨ÈÝÖÁ1L¡£Ã¿´ÎʹÓüÓˮϡÊÍ50±¶¼´1xTAE?buffer¡£
    £¨¶þ£©10?x?TBE?buffer£¨pH8.3£©
    ÔÚ1LÉÕ±­ÖÐ׼ȷ³ÆÈ¡Tris?108g£¬EDTA?7.44g£¬ÅðËá55g£¬¼ÓÈë800mlµÄÈ¥Àë×ÓË®£¬³ä·Ö½Á°è£¬µ÷pH8.3ºó£¬¶¨ÈÝÖÁ1L¡£
    £¨Èý£©6?x?loading?buffer
    ÔÚ500mlÉÕ±­ÖУ¬×¼È·³ÆÈ¡EDTA?4.4g£¬¶þ¼×±½ÇàFF250mg¡¢äå·ÓÀ¶£¨Bromophenol?Blue£©250mg£¬¼ÓÈë200mlÈ¥Àë×ÓË®£¬¼ÓÈÈÖÁ³ä·ÖÈܽ⣻ÔÚ¼ÓÈë180ml¸ÊÓÍ£¬ÌõpHΪ7.0£¬ÓÃÈ¥Àë×ÓË®¶¨ÈÝÖÁ500ml£¬³£Î±£´æ¡£

    ÇíÖ¬ÌÇÄý½ºµçÓ¾½áÊøDNAµÄ»ØÊÕ

    ¶ÔһЩµçÓ¾½áÊøµÄDNA½øÐлØÊÕ£¬ÓÃÓÚÑǿˡ»òÕß̽Õë±ê¼Ç¡£³£Óõķ½·¨ÓеÍÈÛµãÇíÖ¬ÌÇ·¨£¬Ñ¹Ëé·¨¡£»òʹÓÃĿǰ½ÏΪ·½±ãµÄÊÔ¼ÁºÐ»ØÊÕ¡£ÏÂÃæ½éÉܶ³ÈÚ·¨»ØÊÕDNAƬ¶Î¡£

    1. Çнº£¨ÎÞºËËáÎÛȾµÄ¸É¾»µ¶Æ¬ÇУ©£ºµçÓ¾½áÊøºó£¬ÇÐ϶ÔӦƬ¶Î´óСµÄÇíÖ¬ÌÇÄý½º£¬ÖÃÓÚ1.5mlµÄÀëÐĹÜÖУ»
    2. ¼ÓÈëµÈÌå»ýµÄTris-HCl±¥ºÍ·Ó£¨pH7.6£©£¬Õñµ´»ìÔÈ£¬ÔÚ-20¡æ·ÅÖÃ10min£»
    3. µÍÎÂÀëÐÄ£¨10000rpm/5min£©£¬½«ÀëÐÄÉϲã×ªÒÆÖÁеÄÀëÐĹÜÖУ»
    4. ÔÚº¬½ºµÄÀëÐĹÜÖмÓÈëÒ»°ëÌå»ýµÄË®£¬Õñµ´»ìÔÈ£¬ÔÚ-20¡æ·ÅÖÃ10min£»
    5. µÍÎÂÀëÐÄ£¨10000rpm/5min£©£¬ºÏ²¢ÉÏÇåÒº£»
    6. ÓõÈÌå»ýµÄ·ÓºÍÂȷ·ֱð³éÌáÒ»´Î£¬»ñµÃÉÏÇ壻¼ÓÈëÊ®·ÖÖ®Ò»Ìå»ý3M?NaAc£¨pH5.2£©ºÍ2.5±¶Ìå»ýÔ¤ÀäµÄÎÞË®ÒÒ´¼£¬»ìÔÈ£»ÔÚ-20¡æ·ÅÖÃ30min£»
    7. µÍÎÂÀëÐÄ£¨13000rpm/10min£©£¬ÆúÉÏÇ壬75%ÒÒ´¼Ï´³Áµí2´Î£¬´ýÒÒ´¼»Ó·¢ºó¼ÓÈëË®»òÕßTEÈܽ⼴¿É¡£

    Ïà¹Ø·þÎñ

    ¸ü¶àÔĶÁ

    ¼¼Êõ×ÊÔ´

    • µ¥¿Ë¡¿¹ÌåרÌâ
      1. µ¥¿Ë¡¿¹Ìå¸ÅÄî
      2. µ¥¿Ë¡¿¹ÌåÖÆ±¸¼¼Êõ
      3. µ¥Bϸ°û¿¹Ì忪·¢
      4. ¿¹ÌåÑÇÐͼø¶¨
    • ÖØ×鿹ÌåרÌâ
      1. scFv µ¥Á´¿¹Ìå
      2. Ë«ÌØÒìÐÔ¿¹Ìå
      3. ¿¹ÌåÈËÔ´»¯
      4. Ç׺ÍÁ¦³ÉÊì
    • µ°°×±í´ïרÌâ
      1. Ô­ºËµ°°×±í´ï
      2. ²¸È鶯Îïϸ°û±í´ï
      3. ½Íĸµ°°×±í´ï
      4. µ°°×±í´ïFAQ
    • µ°°×´¿»¯×¨Ìâ
      1. His±êÇ©Ç׺ʹ¿»¯
      2. Àë×Ó½»»»É«Æ×´¿»¯
      3. Äý½º¹ýÂËÉ«Æ×·¨
      4. µ°°×´¿»¯±êǩѡÔñ
    • PCR¼¼ÊõרÌâ
      1. ʵʱӫ¹â¶¨Á¿PCR
      2. Äæ×ªÂ¼PCR
      3. RACE¼¼Êõ
      4. ³²Ê½PCR
    • µ°°×»¥×÷¼ì²âרÌâ
      1. ±íÃæµÈÀë×Ó¹²ÕñSPR
      2. ÉúÎïĤ¸ÉÉæBLI
      3. ÃâÒß¹²³ÁµíCo-IP
      4. Èںϵ°°×³Á½µGST Pull down
    ad

    Á¢¿Ìѯ¼Û

    • ÐÕÃû

    • ÁªÏµµç»°

    • ×ÉѯÏîÄ¿

    • ÑéÖ¤Âë

    • ÏîÄ¿ÃèÊö

    Á¢¼´Ñ¯¼Û
    • ÐÕÃû

    • µ¥Î»

    • µç»°

    • ×ÉѯÏîÄ¿

    • ÑéÖ¤Âë

    • ÁôÑÔ:

    ¡¾ÍøÕ¾µØÍ¼¡¿¡¾sitemap¡¿
    ¡¾ÍøÕ¾µØÍ¼¡¿¡¾sitemap¡¿