²úÆ·ÃèÊö
±¾²úÆ·¿ÉÒÔÒÔµ¥Ï¸°û»òÕß΢Á¿×ÜRNAΪģ°å£¬ÔÚͬһ¹ÜÄÚʵÏÖRNAÌáÈ¡¡¢Äæ×ªÂ¼²½Ö裬²»ÐèÒª¶îÍâµÄ²Ù×÷£¬¾ßÓнÚԼʱ¼ä¡¢½µµÍÎÛȾ¡¢Ìá¸ßÁéÃô¶ÈµÈÓÅÊÆ¡£Áí±¾ÊÔ¼ÁºÐÒà¿ÉÊÊÓÃÓÚ1000¸öϸ°ûÒÔϵÄPCRÀ©Ôö£¨Ï¸°ûÓ¦ÔÚPBSÖÐÏ´µÓ£¬ÒòΪÅàÑø»ù»áÓ°ÏìºÏ³ÉcDNAµÄЧÂÊ£©¡£
²úÆ·ÓÅÊÆ
- ¸ßÁéÃô¶È£ºµ¥¸öϸ°û»òµÍÖÁ10pg×ÜRNA¼´¿É×÷ΪÆðʼģ°å£»
- ²Ù×÷±ã½Ý£ºÔÚµ¥¹ÜÄÚʵÏÖÈ«²¿¹¤×÷Á÷³Ì£¬½ÚԼʱ¼ä£¬¼õÉÙÑù±¾×ªÒÆËðʧ£¬½µµÍÎÛȾ£»
- ²úÎïÍêÕûÐԺãºÍ¨¹ýË«¶ËÒýÎïÀ©ÔöÈ«³¤cDNA£¬±ÜÃâÁË5’ºÍ3’Æ«ºÃÐÔ¡£
²úÆ·×é³ÉÓë´¢´æÌõ¼þ£¨96 rxns£©
×é·ÖÃû³Æ |
Ìå»ý |
´¢´æÌõ¼þ |
dNTP Mix |
192¦ÌL |
-20¡æ |
Cell Lysis Buffer |
96¦ÌL |
-20¡æ |
RT Primer |
38.4¦ÌL |
-20¡æ |
DTT |
96¦ÌL |
-20¡æ |
Murine RNase Inhibitor |
38.4¦ÌL |
-20¡æ |
Reverse Transcriptase |
38.4¦ÌL |
-20¡æ |
2 ¡Á PCR Mix |
4mL |
-20¡æ |
PCR1 Primer Mix |
480¦ÌL |
-20¡æ |
RNase-Free water |
2mL |
-20¡æ |
TS Primer |
96¦ÌL |
-80¡æ |
5 ¡Á RT Buffer |
384¦ÌL |
-80¡æ |
Ñù±¾ÀàÐÍ
ϸ°ûÀàÐÍ |
ÊÊÓÃÐÔ˵Ã÷ |
²¸È鶯Îïϸ°û |
°üÀ¨ÈËÔ´¼°ÊóԴϸ°û£¨Èç293T¡¢¸Éϸ°û¡¢ÃâÒßϸ°ûµÈ£©£¬ÐèÈ·±£Ï¸°ûÍêÕûÐÔÓë»îÐÔ¡£ |
Ö²Îï/Õæ¾úϸ°û |
Õë¶Ôϸ°û±Ú½á¹¹ÓÅ»¯ÁѽâÒº£¬ÊÊÓÃÓÚÄâÄϽ桢½ÍĸµÈµ¥Ï¸°û¡£ |
ģʽÉúÎïϸ°û |
Èç¹ûÓ¬¡¢°ßÂíÓã¡¢Ïß³æµÈµÄµ¥Ï¸°û¡£ |
ÌØÊâÀàÐÍϸ°û |
ÂÑϸ°û¡¢Åß̥ϸ°û£¨2-8ϸ°ûÆÚ£©¡¢Ô´úϸ°û£¨ÈçÉñ¾Ôª£©¡¢°©Ï¸°û¼°Ñ»·Ö×Áöϸ°û£¨CTC£©¡£ |
·ÖÑ¡ºóϸ°û |
¼æÈÝÁ÷ʽ·ÖÑ¡£¨FACS£©¡¢Î¢Á÷¿Ø·ÖÑ¡»òÀÂü·ÖÑ¡£¨Èç FlowRACS?£©»ñµÃµÄµ¥Ï¸°ûÐüÒº¡£ |
΢Á¿RNAÑù±¾ |
¿ÉÌæ´úµ¥Ï¸°û£¬Ê¹Óà 10 pg¨C5 ng ×ÜRNAÆðʼ£¨Ï൱ÓÚ1¨C500¸öϸ°û£©¡£ |
×Ô±¸²ÄÁÏ
- ÑùÆ·£ºµ¥Bϸ°ûÑù±¾»ò¶ÔӦ΢Á¿RNA£»
- ÒÇÆ÷£ºÕñµ´Æ÷¡¢Ë²Ê±ÀëÐÄ»ú¡¢PCRÒǵȡ£
×¢ÒâÊÂÏî
- PCR²Ù×÷¾ùÔÚ±ùÉϽøÐУ»
- ËùÓÐPCRÊÔ¼Á¼°ºÄ²ÄÐèÎÞøÎÞ¾ú¡£
ÎïÁ÷ƽ̨£ºÄ¬ÈÏΪ¾©¶«ÎïÁ÷¡£ÈôÓÐÆäËûÐèÇó£¬ÇëÁªÏµ¶Ô½ÓÏúÊÛ¡£
·¢»õʱ¼ä£ºÈ·È϶©µ¥²¢Íê³É¸¶¿îºó1~2¸ö¹¤×÷ÈÕÄÚ°²ÅÅ·¢»õ¡£
·¢»õ·½Ê½£º±ù´ü»ò¸É±ù¡£