浜у搧鍙婄壒鐐更/span>
鑾辨皬鏃犺儐鐢惧師浣撳睘浜庢煍鑶滀綋绾诧紝鏀師浣撶洰锛屾棤鑳嗙斁鍘熶綋绉戯紝鏃犺儐鐢惧師浣撳睘銆侟/span>鑾辨皬鏃犺儐鐢惧師浣撴渶鍒濆垎绂昏嚜姹℃按銆佽偉鏂欍€佽厫鍦熻川銆佸湡澹ゅ拰妞嶇墿缁勭粐锛屽悗鏉ヤ汉浠彂鐜板湪鐗涚敓娈栭亾鍜岄蓟鑵斻€佺尓榧昏厰銆佷汉鍙ｈ厰銆侀浮绐︾瓑鍚勭鍝轰钩鍔ㄧ墿鍜岄笩绫讳腑鍧囧彂鐜板叾瀵勭敓鐜拌薄銆傝幈姘忔棤鑳嗙斁鍘熶綋涔熸槸甯歌鐨勬薄鏌撶粏鑳炵殑鏀師浣撲箣涓€銆傛湰浜у搧鏄?/span>鏍规嵁 PCR鍘熺悊寮€鍙戠殑鑾辨皬鏃犺儐鐢惧師浣撴娴嬭瘯鍓傜洅锛屽畠鍏锋湁涓嬪垪鐗圭偣锛欬/span>

1銆佸嵆寮€鍗崇敤锛岀敤鎴峰彧闇€瑕佹彁渚 DNA妯℃澘銆侟br/>2銆佸紩鐗╃粡杩囩簿蹇冧紭鍖栵紝涓撲竴鎬у己锛屽彧鎵╁鑾辨皬鏃犺儐鐢惧師浣撱€侟br/>3銆佹彁渚涢槼鎬у鐓э紝渚夸簬鍖哄垎鍋囬槾鎬ф牱鍝併€侟br/>4銆丳CR mix涓惈涓婃牱鏌撴枡锛孭CR鍚庡彲浠ョ洿鎺ヤ笂鏍风數娉炽€侟br/>5銆佹湰璇曞墏鐩掕冻澶熷仛 40渭L浣撶郴鐨 PCR 50娆★紝浣嗗彧鑳界敤浜庣鐮斻€侟/span>

瑙勬牸鍙婃垚鍒咟/span>

鎴愪唤	缂栧彿	鍗佸瓟鐩掑寘瑁匋/span>
PCR Magic Mix 3.0	90805	1 mL(绾㈢洊)
瓒呯函姘滁/span>	100935	1 mL(浜粍鑹?
鑾辨皬鏃犺儐鐢惧師浣 PCR寮曠墿娣峰悎娑秉/span>	13-75400yw	100 渭L(鐧界洊)
鑾辨皬鏃犺儐鐢惧師浣 PCR闃虫€у鐓?/span>(1脳108鎷疯礉/渭L)	60908-75400pc	50 渭L(榛勭洊)
浣跨敤鎵嬪唽	13-75400sc	1浠键/span>

杩愯緭鍙婁繚瀛楛/span>
浣庢俯杩愯緭锛?20鈩冧繚瀛橈紝淇濆瓨鏈熼檺涓轰竴骞淬€傞槼鎬у鐓ч渶瑕佸崟鐙斁缃紝涓嶈姹℃煋鍏迭/span>浠栬瘯鍓傘€侟/span>

鑷璇曞墏
鏍峰搧 DNA銆侟/span>

浣跨敤鏂规硶
涓€銆佹牱鍝 DNA鐨勫埗澶嘃/span>

1銆佺敤鑷€夋柟娉曠函鍖 N+2涓牱鍝佺殑 DNA锛屾湰璇曞墏鐩掕窡甯傚満涓婂ぇ澶氭暟鏍搁吀鎻愬彇璇曞墏鐩掑吋瀹广€侟/span>

2銆佸鏋滄湁 N涓牱鍝侊紝鍒欓渶瑕佸仛 N+2涓彁鍙栵紝鍖呮嫭涓€涓牱鍝佸埗澶囬槼鎬у鐓у拰涓€涓牱鍝佸埗澶囬槾鎬у鐓с€傛牱鍝佸埗澶囬槼鎬у鐓ф槸鍦ㄩ€傞噺姘?鍙栧喅浜庤瘯鍓傜洅瑕佹眰鐨勬牱鍝佽捣濮嬮噺)涓姞 10渭L鑾辨皬鏃犺儐鐢惧師浣 PCR闃虫€у鐓х殑 10000鍊嶇█閲婃恫鑰屽緱锛屾牱鍝佸埗澶囬槾鎬у鐓ф槸鐩存帴鐢ㄦ按銆傛彁鍙栫粨鏉熷悗鏈€鍚庡緱鍒版ā鏉 DNA鏀惧啺涓婂緟鐢ㄣ€侟/span>

浜屻€佽缃 PCR鍙嶅簲(40渭L浣撶郴)

3銆佸 N+2涓牱鍝侊紝鍦 PCR鏃堕渶瑕佸鍔犱竴涓 PCR闃虫€у鐓у拰涓€涓 PCR闃存€у鐓э紝鏁呴渶瑕佽缃 N+4涓弽搴斻€傚湪 N+4涓 PCR绠′腑鍒嗗埆鍔犲叆涓嬪垪鎴愬垎锛欬/span>

鎴愬垎	N+2涓牱鍝佺	PCR闃存€у鐓?/span>	PCR闃虫€у鐓?/span>
PCR Magic Mix 3.0	鍚 20 渭L	20 渭L	20 渭L
鑾辨皬鏃犺儐鐢惧師浣 PCR寮曠墿娣峰悎娑秉/span>	鍚 2 渭L	2 渭L	2 渭L
N+2涓牱鍝 DNA妯℃澘	鍚 18 渭L	涓嶅姞	涓嶅姞
PCR闃存€у鐓?姘?	涓嶅姞	18 渭L	涓嶅姞
PCR闃虫€у鐓?鑾辨皬鏃犺儐鐢惧師浣 PCR闃虫€у鐓х殑10000鍊嶇█閲婃恫)	涓嶅姞	涓嶅姞	18 渭L

4銆佹寜涓嬭〃璁剧疆 PCR鍙嶅簲锛欬/span>

杩囩▼	娓╁害	鏃堕棿
棰勫彉鎬?/span>	95鈩傸/span>	5 min
PCR鍙嶅簲(35涓惊鐜?	95鈩傸/span>	30 sec
PCR鍙嶅簲(35涓惊鐜?	55鈩傸/span>	30 sec
PCR鍙嶅簲(35涓惊鐜?	72鈩傸/span>	30 sec
鏈€鍚庡欢浼伏/span>	72鈩傸/span>	10 min

涓夈€佺數娉虫娴婞/span>

5銆佸彇 10-20 渭L PCR浜х墿銆傜數娉虫娴 PCR浜х墿銆傛湰浜у搧鎻愪緵鐨 PCR Mix鍦ㄦ墿澧炵粨鏉熷悗鍙互鐩存帴涓婃牱锛屼笉闇€瑕佸彟澶栧啀鍔 loading buffer銆侾CR浜х墿闃虫€у鐓у繀椤婚鏈熸潯甯﹀嚭鐜帮紝闃存€у鐓у繀椤绘棤浠讳綍鎵╁锛屽惁鍒欏疄楠屾棤鏁堛€傚娌℃湁鎵╁浜х墿鐨勬牱鍝侊紝鍙互绋€閲 10鍊嶅悗閲嶅 PCR鎵╁浠ユ帓闄 PCR鎶戝埗鍓傜殑鎰熸煋銆侟/span>