Northern Blot
鍘熺悊鍙婂疄楠屾柟娉?/p>
鍙戝竷鏃ユ湡锛?/p>
[2012-9-6]
鍏遍槄
[41388]
娆?/p>
鍘熺悊锛?/p>
鍦ㄥ彉鎬ф潯浠朵笅灏嗗緟妫�鐨?/p>
RNA
鏍峰搧杩涜鐞艰剛绯栧嚌鑳剁數娉筹紝
缁ц�屾寜鐓у悓
Southern Blot
鐩稿悓鐨勫師鐞嗚繘琛岃浆鑶滃拰鐢ㄦ帰閽堣繘琛?/p>
鏉備氦妫�娴嬨�?/p>
鐢ㄩ�旓細妫�娴嬫牱鍝佷腑鏄惁鍚湁鍩哄洜鐨勮浆褰曚骇鐗╋紙
mRNA
锛夊強鍏跺惈
閲忋�?/p>
鐐瑰嚮鏌ョ湅鏍搁吀鏉備氦绯诲垪浜у搧
瀹為獙鏂规硶锛?/p>
[
浠櫒銆佽瘯鍓傘�佹潗鏂?/p>
]
锛堜竴锛変华鍣?/p>
鎭掓俯姘存荡绠憋紝鐢垫吵浠紝鍑濊兌鎴愬儚绯荤粺锛岀湡绌鸿浆绉讳华锛岀湡绌烘车锛?/p>
UV
浜よ仈浠紝
鏉備氦鐐夛紝鎭掓俯鎽囧簥锛岃劚鑹叉憞搴婏紝婕╂丁鎸崱鍣紝鍒嗗厜鍏夊害璁★紝寰噺绉绘恫鍣紝鐢?/p>
鐐夛紙鎴栧井娉㈢倝锛夛紝绂诲績绠★紝鐑ф澂锛岄噺绛掞紝涓夎鐡讹紝绛夈�?/p>
锛堜簩锛夋潗鏂?/p>
鎬?/p>
RNA
鏍峰搧鎴?/p>
mRNA
鏍峰搧锛屾帰閽堟ā鏉?/p>
DNA(25 ng)
锛屽凹榫欒啘
锛堜笁锛夎瘯鍓?/p>
NorthernMax Kit
锛?/p>
Cat. # 1940
锛?/p>
Ambion, Inc.
锛?/p>
锛?/p>
鐞艰剛绯栵紝
DEPC, X
鍏夊簳鐗囷紝
搴曠墖鏆楃洅锛?/p>
Random Primer
锛?/p>
dNTP Mixture
锛?/p>
111 TBq/mmol[a-32P]dCTP
锛?/p>
Exo-free Klenow Fragment
鍜?/p>
10 X Buffer, Sephadex G-50,SDS
锛屽弻姘?/p>
姘?/p>
,
鐏弻姘寸瓑銆?/p>
[
鏂规硶涓庢楠?/p>
]
1.
鐢ㄥ叿鐨勫噯澶囷細
180
搴︾儰鍣ㄧ毧锛氫笁瑙掗敟鐡躲�侀噺绛掋�侀晩瀛愩�佸垁鐗囩瓑
4
灏忔椂銆?/p>
鐢垫吵妲斤細娓呮礂姊冲瓙鍜岀數娉虫Ы锛屽苟鐢ㄥ弻姘ф按娴告场杩囧锛岀敤
DEPC
姘村啿娲楋紝骞茬嚗
澶囩敤銆?/p>
澶勭悊
DEPC
姘?/p>
(2 L)
澶囩敤銆?/p>
2
锛庣敤
RNAZaP
鍘婚櫎鐢ㄥ叿琛ㄩ潰鐨?/p>
RNase
閰舵薄鏌?/p>
鐢?/p>
RNAZap
鎿︽礂姊冲瓙锛岀數娉虫Ы锛屽垁鐗囩瓑锛岀劧鍚庣敤
DEPC
姘村啿娲椾簩娆★紝鍘婚櫎
RNAZap
銆?/p>
3
锛庡埗鑳讹細
1
锛?/p>
绉板彇
0.36mg
鐞艰剛绯栧姞鍏ヤ笁瑙掗敟鐡朵腑锛屽姞鍏?/p>
32.4mlDEPC
姘村悗锛屽井娉㈢倝
鍔犵儹鑷崇惣鑴傜硸瀹屽叏鐔旇В銆?/p>
60
鈩冪┖姘旀荡骞宠 婧舵恫
(
闇�鍔?/p>
DEPC
姘磋ˉ鍏呰捀鍙戠殑姘?/p>
鍒?/p>
)
2
锛?/p>
鍦ㄩ�氶鍘ㄤ腑鍔犲叆
3.6ml
鐨?/p>
10
锛?/p>
Denaturing Gel buffer
锛岃交杞绘尟鑽℃贩鍖�銆?/p>
娉ㄦ剰灏介噺閬垮厤浜х敓姘旀场銆?/p>