1.
鐮旂（鍒舵牱锛氬厛鍚戠爺閽典腑鍔犲叆灏戦噺娑叉爱锛岀瓑娑叉爱鎸ュ彂浣跨爺閽甸鍐枫�傚啀鍚戠爺閽靛唴鍊?/p>
鍏ラ�傞噺娑叉爱銆?/p>
浠?/p>
-70
搴﹀啺绠卞彇鍑虹粍缁囷紝
鐢ㄩ晩瀛愪粠鍐诲瓨绠′腑澶瑰彇涓�灏忓潡鏀惧叆鐮旈挼涓�?/p>
杩呴�熺爺纾ㄧ粍缁囧潡锛屾垚绮夋湯鏃跺姞鍏?/p>
1ml
鐨?/p>
trizol
锛岀户缁爺纾紝鍙湅鍒扮矇鏈垚娴嗭紝鐢?/p>
娴撳彉绋�锛岀洿鑷崇粍缁囨牱鍏ㄩ儴婧惰В锛岀敤绉绘恫鏋惛鍒版棤閰?/p>
EP
绠°�?/p>
2. Trizol
娉曟彁鍙?/p>
RNA
1)
瀹ゆ俯闈欑疆
3-10min
锛屼娇铔嬬櫧璐ㄥ彉鎬э紱
2)
鍔犲叆
1/4~1/5
浣撶Н鐨勬隘浠匡紝棰犲�掗渿鑽?/p>
1min
锛屽娓╅潤缃?/p>
3-5min
锛岄潤缃垎灞傦紝鐒?/p>
鍚?/p>
4
鈩冪蹇冿紝
12000rpm
锛?/p>
5min
銆?/p>
锛堟牳铔嬬櫧澶嶅悎浣撳交搴曡瑙ｏ級
3)
鍚稿彇涓婃竻鑷虫棤閰剁殑
EP
绠′腑锛屽姞鍏ョ瓑浣撶Н鐨勯厷锛氭隘浠匡紝棰犲�掗渿鑽?/p>
1min
锛屽娓?/p>
闈欑疆
3-5min
锛岄潤缃垎灞傦紝鐒跺悗
4
鈩冪蹇冿紝
12000rpm
锛?/p>
5min
锛屽彇涓婃竻銆傝嫢鏄粏鑳?/p>
RNA
鎻愬彇鍙彧鍔犵瓑浣撶Н姘豢鎶芥彁涓�娆★紝浣嗗浜庣粍缁囦竴鑸兘瑕侀噸澶嶈姝ラ锛岃嚦娌?/p>
鏈夎泲鐧藉眰涓烘锛?/p>
涓�鑸娊鎻愪袱娆°�?/p>
锛堢蹇冨悗浼氬垎涓轰笁娆★細
涓婂眰锛?/p>
姘寸浉锛?/p>
涓棿灞傦細
DNA
锛?/p>
涓嬪眰锛?/p>
铔嬬櫧銆?/p>
涓轰簡闄嶄綆姘寸浉鍜屾湁鏈虹浉鍒嗙晫澶?/p>
DNA
姹℃煋锛?/p>
涓嶈鍚稿彇姘寸浉鐨勬渶涓嬪眰锛?/p>
4)
鍔犲叆绛変綋绉殑棰勫喎鐨勫紓涓欓唶锛岄鍊掓贩鍖�锛屽啺涓婇潤缃?/p>
30min
锛岀劧鍚?/p>
4
鈩冪蹇?/p>
12000rpm
锛?/p>
30min
锛屽純涓婃竻銆?/p>
锛堟姝ラ涓昏鏄厖鍒嗘矇娣�
RNA
锛岀劧鍚庢敹闆嗐�傛晠姝ラ
涓殑鍐扮疆
30min,
涔熷彲浠ユ槸鍦?/p>
-20掳
鏀剧疆鍗婂皬鏃朵互涓婏紝鐢氳嚦杩囧锛?/p>
5)
鍔犲叆
100ul 70%
涔欓唶锛?/p>
灏嗘矇娣�鍚硅捣鏉ワ紝
娉ㄦ剰涓嶈鍚规暎锛?/p>
鐒跺悗
4
鈩?/p>
12000rpm
锛?/p>
5min
銆?/p>
6)
寮冧笂娓咃紝绌烘皵涓尌鍙戜箼閱囷紝鑷冲涓婃棤娑叉淮銆?/p>
7)
鍔犲叆閫傞噺
DEPC
姘达紝
锛堜竴鑸负
20ul
锛夋憾瑙ｃ�?/p>
8)
鍒嗗厜鍏夊害璁℃娴?/p>
RNA
娴撳害鍜?/p>
OD
鍊硷紝灏?/p>
Total RNA
绋�閲婅嚦
1ug/ul
锛屽鐢ㄣ�?/p>