缁嗚弻鍩哄洜缁?/p>
DNA
鎻愬彇璇曞墏鐩掞紙绂诲績鏌卞瀷锛?/p>
GK4003-10
GK4003-20
GK4003-50
GK4003-100
涓�
.
璇曞墏鐩掔粍鎴?/p>
Components
GK4003-10
10 Preps
GK4003-20
20 Preps
GK4003-50
50 Preps
GK4003-100
100 Preps
GenClean Column
2.0-ml Collection Tube
Lysis Solution
AW1 Solution
(a)
AW2 Solution
(b)
AE Buffer
(c)
TE(pH8.0)
Boiled RNase A
Proteinase K
(d)
Lysozyme
(f)
SP Buffer
protocol
10
10
4 ml
6 ml
3 ml
3 ml
2 ml
50 碌
l
30 碌
l
5 mg
1.0 ml
1
20
20
10 ml
12 ml
8 ml
8 ml
4 ml
240 碌
l
250 碌
l
15 mg
1.0 ml
1
50
50
25 ml
30 ml
20 ml
20 ml
10 ml
480 碌
l
500 碌
l
30 mg
1.0 ml
1
100
100
50 ml
60 ml
40 ml
40 ml
20 ml
120 碌
l
1000 碌
l
60 mg
1.0 ml
1
(a)
棣栨浣跨敤鍓嶏紝蹇呴』鍦?/p>
AW1
婧舵恫鐡朵腑鎸夎〃绀洪噺鍔犲叆鏃犳按涔欓唶锛屽厖鍒嗘贩鍖�鍚庝娇鐢ㄣ�傛瘡娆′娇鐢ㄥ悗灏嗙摱鐩栫洊
绱э紝浠ヤ繚鎸佷腑鐨?/p>
AW1
涔欓唶鍚噺銆傚鏋滄偍鍙戠幇
AW1
鐢变簬杩愯緭鎴栦繚绠′笉褰撻�犳垚瀹归噺涓ラ噸涓嶅噯锛岃鐢ㄩ噺绛掑畾
瀹瑰悗鍐嶅姞鍏?/p>
0.6
鍊嶄綋绉殑鏃犳按涔欓唶銆?/p>
(b)
棣栨浣跨敤鍓嶏紝蹇呴』鍦?/p>
AW2
鐡朵腑鍔犲叆
4
鍊嶄綋绉殑鏃犳按涔欓唶锛屽厖鍒嗘贩鍖�鍚庝娇鐢ㄣ�傛瘡娆′娇鐢ㄥ悗灏嗙摱鐩栫洊绱э紝
浠ヤ繚鎸?/p>
AW2
涓殑涔欓唶鍚噺銆傚鏋滄偍鍙戠幇
AW2
鐢变簬杩愯緭鎴栦繚绠′笉褰撻�犳垚瀹归噺涓ラ噸涓嶅噯锛岃鐢ㄩ噺绛掑畾瀹瑰悗
鍐嶅姞鍏?/p>
4
鍊嶄綋绉殑鏃犳按涔欓唶銆?/p>
(d) AE Buffer
涓?/p>
10 mM Tris-HCl, 0.5 mM EDTA pH 8.5
銆?/p>
TE(pH 8.0)
鎴栬�呮按锛?/p>
pH>7.0
锛夊潎鍙互浣跨敤锛屼絾
鏄?/p>
DNA
鐨勬礂鑴辨晥鐜囬�氬父瑕佷綆
20%
宸﹀彸銆?/p>
(e)
鏀跺埌鍚庡垎瑁?/p>
-20
鈩冨喕瀛樸�備笉寰楀皢
Proteinase K
鐩存帴鍔犲埌
Lysis Solution
涓紝浠ュ厤閰跺け娲汇�?/p>
(f) Lysozyme
浣跨敤鍓嶅姞鍏ユ寜
5 mg
婧朵簬
100碌
l
璁＄畻锛屾憾浜?/p>
SP Buffer
锛屾憾瑙ｅ悗锛屽垎瑁呬簬
-20
鈩冨喕瀛樸�?/p>
瀹㈡埛鑷锛?/p>
PBS
缂撳啿娑诧紝鑳拌泲鐧介叾锛屾棤姘翠箼閱?/p>
杩愯緭
鍌ㄥ瓨娓╁害
杩愯緭
瀹ゆ俯
鍌ㄥ瓨
Proteinase K , Lysozyme , Boiled RNase A :
-20
鈩?/p>
鍏朵粬
瀹ゆ俯
浜岋紟涓昏鐢ㄩ�?/p>
浠庣粏鑿屽拰鍚勭鍩瑰吇缁嗚優涓皬瑙勬ā鎶芥彁鍩哄洜缁?/p>
DNA
銆?/p>
涓夛紟涓昏鐗圭偣
1.
绂诲績鍚搁檮鏌卞唴纭呭熀璐ㄨ啘鍏ㄩ儴閲囩敤杩涘彛鐗瑰埗鍚搁檮鑶滐紝鏌变笌鏌变箣闂村惛闄勯噺宸紓鏋佸皬锛岄噸澶嶆�уソ銆?/p>
2.
涓嶉渶瑕佷娇鐢ㄦ湁姣掔殑鑻厷绛夎瘯鍓傦紝涔熶笉闇�瑕佷箼閱囨矇娣�绛夋楠ゃ�?/p>
3.
蹇�燂紝绠�鎹凤紝鍗曚釜鏍峰搧鎿嶄綔涓�鑸彲鍦?/p>
50
鍒嗛挓鍐呭畬鎴愩�?/p>
4.
澶氭鏌辨紓娲楃‘淇濇彁鍙栫粏鑿屽熀鍥犵粍鐨勯珮绾害锛?/p>
OD
260/280nm
鍏稿瀷鐨勬瘮鍊艰揪
1.7
锝?/p>
1.9
锛岄暱搴﹀彲杈?/p>
50
锝?/p>
100kb
锛屽彲
鐩存帴鐢ㄤ簬
PCR
锛?/p>
Southern-blot
鍜屽悇绉嶉叾鍒囧弽搴斻�?/p>
鍥涳紟鍩瑰吇缁嗚優涓庣粏鑿屾牱鍝佸噯澶?/p>
1
锛庡煿鍏荤粏鑳炵殑鍑嗗
(1)
瀵逛簬鎮诞鍩瑰吇缁嗚優锛?/p>
1,000g
瀹ゆ俯绂诲績
5
鍒嗛挓锛屽交搴曞幓闄や笂娓呫�?/p>