AFLP
鎶�鏈殑鍩烘湰鍘熺悊涓庡疄楠屾柟娉?/p>
AFLP
鎶�鏈殑鍩烘湰鍘熺悊
:
AFLP
鎶�鏈槸涓�椤规柊鐨勫垎瀛愭爣璁版妧鏈紝
鍏跺師鐞嗘槸锛?/p>
鍩哄洜缁?/p>
DNA
缁忚繃浜岀閰朵笉鍚岀殑闄愬埗鎬у唴鍒囬叾閰跺垏鍚庯紝
浜х敓绮?/p>
鎬ф湯绔紝
鍐嶄娇鐢ㄨ繛鎺ラ叾灏嗕汉宸ュ悎鎴愮殑鍙岄摼鎺ュご杩炴帴鍦ㄩ叾鍒囦綅鐐圭殑绮樻�ф湯绔�?/p>
鎺ュご涓�绔叿鏈変笌鍐呭垏閰跺悓鏍风殑璇嗗埆绮?/p>
鎬ф湯绔紝浜掕ˉ杩炴帴鍚庢垚涓?/p>
DNA
妯℃澘杩涜棰勬墿澧炪�傛帴澶村拰涓庢帴澶寸浉閭荤殑閰跺垏鐗囨柇鐨勫嚑涓⒈鍩哄簭鍒椾綔涓哄紩鐗╃殑缁撳悎浣?/p>
鐐广�傚紩鐗╃敱
3
閮ㄥ垎缁勬垚
:
鈶犳牳蹇冪⒈鍩哄簭鍒楋紝璇ョ⒈鍩哄簭鍒椾笌浜哄伐鎺ュご浜掕ˉ锛涒憽鐗瑰紓鎬ч叾鍒囧簭鍒楋紱鈶㈠紩鐗?/p>
3
鈥欑閫夋嫨鎬?/p>
纰卞熀銆?/p>
閫夋嫨鎬х⒈鍩哄欢浼稿埌閰跺垏鐗囨鍖猴紝
杩欐牱灏卞彧鏈夐偅浜涗袱绔簭鍒楄兘涓庨�夋嫨纰卞熀閰嶅鐨勯檺鍒舵�ч叾鍒囩墖娈佃鎵╁銆?/p>
鍙?/p>
澶栵紝
閫氳繃閫夋嫨鍦ㄦ湯绔垎鍒坊鍔犱簡
1
锝?/p>
3
涓�夋嫨鎬ф牳鑻烽吀鐨勪笉鍚屽紩鐗╋紝
鍙互杈惧埌閫夋嫨鎵╁鐨勭洰鐨勩�?/p>
杩欎簺閫夋嫨鎬ф牳鑻烽吀
浣垮緱寮曠墿閫夋嫨鎬у湴璇嗗埆鍏锋湁鐗瑰紓閰嶅搴忓垪鐨勫唴鍒囬叾閰跺垏鐗囨銆傚苟涓庝箣缁撳悎锛屽疄鐜扮壒寮傛�ф墿澧炪�?/p>
瀹為獙鏂规硶
:
(1)
DNA
閰跺垏
AFLP
鎶�鏈垚鍔熺殑鍏抽敭鍦ㄤ簬
DNA
鐨勫厖鍒嗛叾鍒?/p>
,
鎵�浠ュ妯℃澘璐ㄩ噺瑕佹眰杈冮珮
,
鍦?/p>
DNA
瀹屽叏婧惰В鍚庡埄鐢ㄧ传澶栧垎鍏夊厜搴︿华
娴嬪畾
DNA
娴撳害锛屽皢
DNA
娴撳害鐢ㄥ弻钂告按璋冩暣鍒?/p>
50ng/ul,
搴旈伩鍏嶅叾浠?/p>
DNA
姹℃煋鍜屾姂鍒剁墿璐ㄧ殑瀛樺湪銆?/p>
琛?/p>
1
锛?/p>
E-M
閰跺垏浣撶郴
鍙嶅簲浣撶郴锛?/p>
E-M
缁?/p>
鍚堬級
1/2
鍊?/p>
1
鍊?/p>
DNA
锛?/p>
50ng/ul
锛?/p>
1.50ul
3.0 ul
EcoR
鈪?/p>
(10u/ ul)
0.25ul
0.5 ul
Mse
鈪?/p>
(10u/ ul)
0.15ul
0.3 ul
Tango
Buffer (10
脳
)
2.50ul
5.0 ul
ddH
2
O
8.10ul
16.2
ul
鎬讳綋绉?/p>
12.5ul
25 ul
琛?/p>
2
锛?/p>
P-M
閰跺垏浣撶郴
鍙嶅簲浣撶郴锛?/p>
P-M
缁?/p>
鍚堬級
1/2
鍊?/p>
1
鍊?/p>
DNA
锛?/p>
50ng/ul
锛?/p>
1.50ul
3.0 ul
Pst
鈪?/p>
(10u/ ul)
0.25ul
0.5 ul
Mse
鈪?/p>
(10u/ ul)
0.15ul
0.3 ul
Buffer R (10
脳
)
1.25ul
2.5 ul
ddH
2
O
9.35ul
18.7
ul
鎬讳綋绉?/p>
12.5ul
25 ul