ÊɾúÌåչʾÎÄ¿âµÄ¹¹½¨Óëɸѡ

ÊɾúÌåչʾ¼¼ÊõÊÇÒ»ÖÖÓÃÓÚʶ±ðµ°°×ÖʺÍÆäËû´ó·Ö×ÓÅäÌåµÄÌåÍâɸѡ¼¼Êõ¡£ÓÉSimthÓÚ1985Ä꽨Á¢¡£ÆäÔ­ÀíÊǽ«ÍâÔ´¶àëÄ»òµ°°×ÖʵÄDNAÐòÁвåÈëµ½ÊɾúÌåÍâ¿Çµ°°×µÄÒ»¸ö½á¹¹»ùÒòµÄÊʵ±Î»ÖÃÉÏ£¬ÔÚÔĶÁ¿òÕý³£ÇÒ²»Ó°ÏìÍâ¿Çµ°°×Õý³£¹¦ÄܵÄÇé¿öÏ£¬Ê¹ÍâÔ´»ùÒòËæ×ÅÍâ¿Çµ°°×µÄ±í´ï¶ø±í´ï£¬×îÖÕ¶àëÄ»òµ°°×ÒÔÓëÍâ¿Çµ°°×ÈںϵÄÐÎʽչʾÔÚÊɾúÌå±íÃæ¡£±»Õ¹Ê¾µÄµ°°×»òÕ߶àëÄ¿ÉÒÔ±£³ÖÏà¶ÔµÄ¿Õ¼ä½á¹¹ºÍÉúÎï»îÐÔ£¬¿ÉÒÔÀûÓðе°°×¶ÔÆä½øÐÐɸѡ¡£

¸ßÇ׺ÍÁ¦¿¹Ìåɸѡ

ÊɾúÌ忹Ìå¿â¹¹½¨

ÀûÓÃÊɾúÌåչʾ¼¼Êõ½øÐиßÇ׺ÍÁ¦¿¹ÌåµÄɸÑ¡£¬ÐèÒªÏȹ¹½¨¿¹Ìå¿â£¬È»ºóÔÙ¶ÔÊɾúÌåÎÄ¿â½øÐÐɸѡ¡£½«ÍâÔ´»ùÒò²åÈëÊɾúÌå»ùÒòÊʵ±Î»Öã¬Ëæ×ÅÊɾúÌåµÄ´«´ú£¬ÍâÔ´µ°°×Õ¹ÏÖÔÚ×Ó´úÊɾúÌåµÄ±íÃæ¡£ËùÓÐչʾ²»Í¬ÍâÔ´»ùÒòµÄÊɾúÌåµÄ¼¯ºÏ£¬¼´ÊɾúÌåչʾÎÄ¿â¡£

ÓÃÓÚ¹¹½¨ÎÄ¿âµÄ²»Í¬ÐòÁеĺËËáƬ¶ÎÓÐÈ˹¤ºÏ³É·¨¡¢cDNA·¨¡¢DNase IËæ»úË®½âDNAµÈ·½·¨»ñµÃ¡£ÓÃÓÚ¹¹½¨ÎÄ¿âµÄÊɾúÌåչʾϵͳÓе¥Á´Ë¿×´ÊɾúÌåչʾϵͳ¡¢¦ËÊɾúÌåչʾϵͳ¡¢T4ÊɾúÌåչʾϵͳ¼°T7ÊɾúÌåչʾϵͳµÈ¡£
ÀûÓÃcDNA·¨»ñµÃ»ùÒòƬ¶Î²¢½øÐÐÊɾúÌåչʾÎÄ¿â¹¹½¨µÄ²Ù×÷Á÷³ÌΪÌáÈ¡mRNA £»Äæת¼µÃµ½cDNA £»Éè¼ÆºÏÊÊÒýÎÒÔcDNAΪģ°å£¬PCRµÃµ½»ùÒòÐòÁУ»½«»ùÒò¿Ë¡ÖÁÊɾúÌåÔØÌåÖУ»µçת¸ÐÊÜ̬ϸ°û£¬¸¨ÖúÊɾúÌ峬Ⱦ£»ÊÕ¼¯ÉÏÇ壬ÉÏÇ弴ΪÊɾúÌåչʾÎÄ¿â¡£

ͼ1£ºÊɾúÌåչʾÎÄ¿â¹¹½¨Á÷³Ì

ÊɾúÌ忹Ìå¿âµÄɸѡ

ɸѡµÄ·½·¨Ö÷Òª·ÖΪÌåÄÚɸѡºÍÌåÍâɸѡÁ½ÖÖ¡£ÌåÄÚɸѡÊÇÖ¸½«ÊɾúÌåչʾÎÄ¿â¾²Âö×¢Éäµ½¶¯ÎïÌåÄÚ£¬ÒòΪѪ¹Ü·Ö×ÓÄÚƤµÄÒìÖÊÐÔ£¬ÊɾúÌå¿ÉÒÔÓÐÑ¡ÔñÐԵĵ¼Ïò²»Í¬×éÖ¯£¬ÕâÑù¾Í¿ÉÒԵõ½Ó벻ͬ×éÖ¯ÌØÒì½áºÏµÄÊɾúÌåչʾ¿¹Ì壻ÌåÍâɸѡµÄ·½·¨ÓÐÉúÎïÌÔÏ´·¨¡¢¾ºÕù·¨¡¢Ïû¼õ·¨¡¢Ñ¡ÔñÐÔ¸ÐȾɸѡ¡¢ÑÓ³ÙÐÔ¸ÐȾɸѡµÈ·½·¨¡£ÆäÖÐÉúÎïÌÔÏ´·¨×îΪ³£Óᣱ¾ÎÄÖ÷Òª¶ÔÉúÎïÌÔÏ´·¨É¸Ñ¡ÊɾúÌ忹Ìå¿â½øÐÐ˵Ã÷¡£

ÉúÎïÌÔÏ´·¨£ºÒ԰е°°×Ϊ¹Ì¶¨Ï࣬ÒÔÊɾúÌåչʾÎÄ¿âΪÁ÷¶¯Ï࣬¾­¹ýÒ»¶Îʱ¼äµÄ¹²Í¬·õÓýºó£¬Ï´È¥Î´½áºÏµÄÓÎÀëÊɾúÌ壬ȻºóÒÔ¾ºÕùÊÜÌå»òËáÏ´ÍÑÏÂÓë°Ð·Ö×Ó½áºÏÎü¸½µÄÊɾúÌ壬ϴÍѵÄÊɾúÌå¸ÐȾËÞÖ÷ϸ°ûºó¾­·±Ö³À©Ôö£¬½øÐÐÏÂÒ»ÂÖÏ´ÍÑ£¬¾­¹ý3ÂÖ¡«5Âֵġ°Îü¸½-Ï´ÍÑ-À©Ôö¡±ºó£¬¼´¿ÉµÃµ½Óë°Ðµ°°×¾ßÓиßÇ׺ÍÐԵĿ¹Ìå¡£

ͼ2£ºÊɾúÌåչʾ¼¼ÊõɸѡÁ÷³Ì

¾­¹ýɸѡºó£¬¶ÔɸѡµÃµ½µÄµ°°×½øÒ»²½ÓÃøÁªÃâÒßÎü¸½²â¶¨»òÃâÒßÓ¡¼£·¨¶ÔÆä½øÐÐÌØÒìÐÔɸÑ¡£¬»ñÈ¡Ä¿µÄ¿Ë¡»òÆÌÅ̽øÐе¥¿Ë¡·ÖÀë¼Ì¶ø½øÐдó¹æÄ£Éú²ú¡£

Ïà¹ØÔĶÁ

ÊɾúÁ£ÓëÊɾúÁ£ÔØÌå
PCR¼¼Êõ