Protocol Online logo
Top : Forum Archives: : Methods Discussion

ÇóÖúMSP - ±ØÐëøÇйýÒ¹Âð (Feb/01/2005 )

ÎÒÕýÔÚÓÃÈ˵ĻùÒò×é×öMSP
1.±ØÐëøÇйýÒ¹Âð?
2.ÄÜ°ïÎÒ¿´Ò»ÏÂʵÑé²½ÖèÂð£¿
È¡»ùÒò×éDNA10 ul£¬Å¨¶ÈԼΪ100-200ng/ul£¬¼Ó40ulÃð¾ú´¿Ë®£¬»ìÔÈ¡£
¼ÓÐÂÏÊÅäÖÃ3MÇâÑõ»¯ÄÆÈÜÒº5 ul£¬42¡æ±£ÎÂ30·ÖÖÓ¡£
ÆÚ¼äÅäÖÆ40.5%ÑÇÁòËáÇâÄƺÍ10mMÇâõ«£¬ÆäÖÐÑÇÁòËáÇâÄÆÓÃ3MÇâÑõ»¯ÄƵ÷½ÚPHÖµÖÁ5¡£
10mMÇâõ«
40.5%ÑÇÁòËáÇâÄÆ 8.1g ÑÇÁòËáÇâÄÆ
17ml Ë®
10N ÇâÑõ»¯ÄÆ µ÷pHµ½5.0
¼ÓË®ÖÁ20 ml
È¡³öÑùÆ·£¬¼ÓÈë10mMÇâõ«30ul£¨ÈÜÒºÑÕɫת»Æ£©£¬
¼ÓÈë40.5%ÑÇÁòËáÇâÄÆ510 ul£¬Îý²­Ö½°ü¹üÑùÆ·¡£
ÇáÈá»ìÔÈ£¬55¡æ±£ÎÂ16h¡£
ÓÃDNA clean-upÊÔ¼ÁºÐ´¿»¯DNA£¬ÊÕ¼¯50ulÏ´ÍÑÒº¡£
¼Ó5.5ulŨ¶ÈΪ3NµÄÇâÑõ»¯ÄÆ£¬»ìÔÈ£¬37¡æ·ÅÖÃ15·ÖÖÓ¡£
È»ºóÔÙ¼Ó8MÒÒËá泥¨pH7.0)ÈÜÒº35ul£¬»ìÔÈ¡£
¼ÓÔ¤ÀäÎÞË®ÒÒ´¼300ul£¬µßµ¹»ìÔÈ£¬ÖÃ-20¡æ±ùÏä¹ýÒ¹¡£
È¡³öÑù±¾£¬13200rpmÀëÐÄ20min£¬ÆúÈ¥ÉÏÇ壬¼Ó70%ÒÒ´¼100ul£¬13200rpmÀëÐÄ2min£¬ÆúÈ¥ÉÏÇ壬ÁÀ¸ÉDNA£¬¼ÓÈëÃð¾ú´¿Ë®30ul¡£

-ÐÂÊÖÇóÖú-

ÄãµÄʵÑé²½ÖèûÓÐʲôÎÊÌâ¡£

¹ØÓÚøÇеÄÎÊÌ⣬²»Ò»¶¨ÊDZØÐëµÄ£¬Èç¹ûÓÐDNAת»»²»ÍêÈ«µÄÇé¿ö£¬ÓÃøÇÐһϻáÓаïÖú¡£Ö
»ÓÐËùÓõÄø²»ÇÐÄãµÄÀ©ÔöƬ¶Ï£¬Ê²Ã´Ã¸¶¼¿ÉÒÔ¡£

-hula-

QUOTE (hula @ Feb 1 2005, 10:36 AM)
ÄãµÄʵÑé²½ÖèûÓÐʲôÎÊÌâ¡£

¹ØÓÚøÇеÄÎÊÌ⣬²»Ò»¶¨ÊDZØÐëµÄ£¬Èç¹ûÓÐDNAת»»²»ÍêÈ«µÄÇé¿ö£¬ÓÃøÇÐһϻáÓаïÖú¡£Ö
»ÓÐËùÓõÄø²»ÇÐÄãµÄÀ©ÔöƬ¶Ï£¬Ê²Ã´Ã¸¶¼¿ÉÒÔ¡£

żÍêÈ«Õâ¸ö²½Ö裬һÇж¼·Ç³£ok£¬NaOH͵ÀÁÒ²²»ÊÇÿ´ÎÐÂÏÊÅ䣬 rolleyes.gif ½á¹ûºÜ²»´íµÄhappy.gif

-Rovy-