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shelly9018

新虫 (小有名气)

[求助] Southern blot探针标记不理想,但标记效率检测时,对照显色也不理想?

如题,新手做Southern blot探针标记,用质粒酶切回收片段作为模板,但一直回收的浓度都不高,最高的这次是54ng/ul,取了10ul按罗氏试剂盒操作步骤做探针,然后按试剂盒说明做标记效率检测,结果如图所示

经过过夜显色(16h),对照只出来三个浓度(10pg、3pg、1pg),而且颜色都很淡,所呈现的斑点颜色还不均匀,按试剂盒的说明,显色这么长时间,至少0.1pg的显示都应该出来了,新制备的探针值只出现了理论最高浓度(10pg),如果说探针的标记不理想的话,主要的原因会是什么? 对照的显色也不理想,这个原因一直没有想明白???请各位虫友多多帮忙,江湖救急啊

Southern blot探针标记不理想,但标记效率检测时,对照显色也不理想?
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