F**********6 发帖数: 90 | 1 做了几次CHIP,每次对照IgG都能扩增出很弱的条带(实验组的条带很强!)。请问如
何解决对照IgG有弱条带的问题?
看了些别人发表的CHIP实验数据,绝大部份对照IgG基本上没有条带。但偶然能看到一
两篇文章里的CHIP实验数据里的对照里有弱的条带。
多谢指教 | m********m 发帖数: 263 | 2 ChIP只是反映你的目的条带的enrichment,IgG出现很弱的条带没有关系,如果想去除
,可以使用高盐buffer多洗几次。洗的次数得根据结合强弱来判定。
【在 F**********6 的大作中提到】![](/moin_static193/solenoid/img/up.png) : 做了几次CHIP,每次对照IgG都能扩增出很弱的条带(实验组的条带很强!)。请问如 : 何解决对照IgG有弱条带的问题? : 看了些别人发表的CHIP实验数据,绝大部份对照IgG基本上没有条带。但偶然能看到一 : 两篇文章里的CHIP实验数据里的对照里有弱的条带。 : 多谢指教
| d*****s 发帖数: 647 | | F**********6 发帖数: 90 | | A******y 发帖数: 2041 | 5 If you use IgG as control for CHIP, some reviewers will kill you saying you
didn't do the proper control. Find a transcription factor that is known NOT
to bind to the region and use it as a negative control.
Of course, if you see the non-specific band, expose less or wash harder. However, CHIP is level of enhancement very different from IP for protein. | Z******5 发帖数: 435 | 6 一般都是IgG做Control吧,貌似还没见过拿其它转录因子做Control的。
you
NOT
However, CHIP is level of enhancement very different from IP for protein.
【在 A******y 的大作中提到】![](/moin_static193/solenoid/img/up.png) : If you use IgG as control for CHIP, some reviewers will kill you saying you : didn't do the proper control. Find a transcription factor that is known NOT : to bind to the region and use it as a negative control. : Of course, if you see the non-specific band, expose less or wash harder. However, CHIP is level of enhancement very different from IP for protein.
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