涓撻
09
鐜颁唬鐢熺墿鎶�鏈�夊仛棰?/p>
鐜颁唬鐢熺墿鎶�鏈紝鏄敓鐗╃瀛︿笌宸ョ▼鎶�鏈湁鏈虹粨鍚堣�屽叴璧风殑涓�闂ㄧ患鍚堟�х瀛︽妧鏈紝鍖呮嫭鍩哄洜宸ョ▼銆佺粏鑳?/p>
宸ョ▼銆佽儦鑳庡伐绋嬬瓑涓�绯诲垪鐨勭幇浠ｇ敓鐗╂妧鏈紝鍏充簬鍩哄洜宸ョ▼銆佺粏鑳炲伐绋嬨�佽儦鑳庡伐绋嬬殑鏉愭枡棰樻槸杩戝勾鏉ラ珮鑰冪殑
鐑棬鐭ヨ瘑鐐广�傛湰閮ㄥ垎灞炰簬楂樿�冪殑閲嶇偣鍐呭涔嬩竴锛屽涔犳椂瑕侀噸瑙嗙敓鐗╂柊鎶�鏈湪鐢熶骇鐢熸椿瀹炶返涓殑搴旂敤锛岄噸瑙?/p>
鍚勫伐绋嬩箣闂寸殑鐩镐簰鍏崇郴锛岃繖鏃㈡槸绉戞妧鍙戝睍鐑偣璇鹃涔嬩竴锛屼篃鏄�冭瘯鍛介鐑偣涔嬩竴銆傚埄鐢ㄥ凡瀛﹁繃鐨勫熀鍥犲伐绋嬨�?/p>
缁嗚優宸ョ▼銆佽儦鑳庡伐绋嬬煡璇嗭紝瀵逛笁澶х敓鐗╁伐绋嬬殑鑱旂郴鍔犱互铻嶄細璐�氾紝鎻愰珮鍒嗘瀽缁煎悎涓庤В鍐冲疄闄呴棶棰樼殑鑳藉姏銆?/p>
1.RT
锛?/p>
PCR
鏄皢
RNA
閫嗚浆褰?/p>
(RT)
鍜?/p>
cDNA
鐨勮仛鍚堥叾閾惧紡鎵╁鍙嶅簲鐩哥粨鍚堢殑鎶�鏈紝鍏蜂綋杩囩▼濡傚浘鎵�绀猴細
(1)
杩囩▼
鈪?/p>
闇�瑕佸姞鍏ョ紦鍐叉恫銆佸師鏂欍�?/p>
_____________
銆?/p>
_______________
鍜屽紩鐗?/p>
A
绛夈�?/p>
(2)
杩囩▼
鈪?/p>
棣栧厛瑕佸皢鍙嶅簲浣撶郴鐨勬俯搴﹀崌楂樺埌
95
鈩?/p>
锛屽叾鐩殑鏄?/p>
__________________
銆?/p>
_______________
銆?/p>
(3)
杩囩▼
鈪?/p>
鎷熷鍗曢摼
cDNA
杩涜
n
娆″惊鐜殑鎵╁锛岀悊璁轰笂鑷冲皯闇�瑕?/p>
_________
涓紩鐗?/p>
B
銆?/p>
(4)
鍒╃敤
RT
锛?/p>
PCR
鎶�鏈幏鍙栫殑鐩殑鍩哄洜
_______________ (
濉?/p>
鈥?/p>
鑳?/p>
鈥?/p>
鎴?/p>
鈥?/p>
涓嶈兘
鈥?
鍦ㄧ墿绉嶄箣闂翠氦娴侊紱璇ユ妧鏈繕鍙敤
浜庡鏌愪簺寰噺
RNA
鐥呮瘨鐨勬娴嬶紝鍙彁楂樻娴嬬殑鐏垫晱搴︼紝鍘熷洜鏄?/p>
_______
銆?/p>
(5)RT
锛?/p>
PCR
杩囩▼涓富瑕佸�熷姪瀵?/p>
__________
鐨勬帶鍒讹紝褰卞搷閰剁殑娲绘�э紝浠庤�屼娇寰楀寲瀛﹀弽搴旀湁搴忛珮鏁堝湴杩涜銆?/p>
銆愮瓟妗堛�?/p>
(1).
RNA
鎻愬彇鐗?/p>
(2).
閫嗚浆褰曢叾
(3).
璁╅�嗚浆褰曢叾鍙樻�уけ娲?/p>
(4).
浣?/p>
mRNA
锛?/p>
cDNA
鏉傚悎
鍙岄摼瑙ｅ紑
(5).
2
n
锛?/p>
1
(6).
鑳?/p>
(7).
澧炲姞浜嗗緟娴?/p>
RNA
閫嗚浆褰曚骇鐢熺殑
DNA
鐨勬暟閲?/p>
(
鎴栨祿搴?/p>
)
锛屼究浜庢娴?/p>
(8).
娓╁害
銆愯В鏋愩�?/p>
(1)
杩囩▼
鈪?/p>
浠?/p>
mRNA
涓烘ā鏉垮悎鎴?/p>
cDNA
鐨勮繃绋嬶紝琛ㄧず閫嗚浆褰曪紝闇�瑕佸姞鍏ョ紦鍐叉恫銆佸師鏂欍�?/p>
RNA
鎻愬彇
鐗┿�侀�嗚浆褰曢叾鍜屽紩鐗?/p>
A
绛夈�?/p>
(2)
杩囩▼
鈪?/p>
浠?/p>
mRNA
涓烘ā鏉垮悎鎴?/p>
cDNA
锛?/p>
褰㈡垚
cD
NA-mRNA
鏉傚悎鍙岄摼銆?/p>
鍥犳锛?/p>
杩囩▼
鈪?/p>
棣栧厛瑕佸皢鍙嶅簲浣撶郴鐨勬俯
搴﹀崌楂樺埌
95
鈩?/p>
锛屼娇
mRNA
锛?/p>
cDNA
鏉傚悎鍙岄摼瑙ｅ紑锛屽悓鏃惰閫嗚浆褰曢叾鍙樻�уけ娲汇�?/p>
(3)
鏍规嵁
DNA
鍒嗗瓙鍗婁繚鐣欏鍒剁殑鐗圭偣锛屽鍗曢摼
cDNA
杩涜
n
娆″惊鐜殑鎵╁锛岀悊璁轰笂鑷冲皯闇�瑕?/p>
2
n-1
涓紩鐗?/p>
B
銆?/p>
(4)
涓嶅悓鐢熺墿浣撳唴
DNA
鍒嗗瓙鐨勫熀鏈粨鏋勭浉鍚岋紝涓旀墍鏈夌殑鐢熺墿鍏辩敤涓�濂楀瘑鐮佸瓙锛屽洜姝ゅ埄鐢?/p>
RT
锛?/p>
PCR
鎶�鏈幏鍙?