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DNA ladders on a northern blot ?..


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#1 alexsunny123

alexsunny123

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Posted 18 December 2020 - 01:38 AM

Hello,

 

I was wondering if someone could tell me if this would work;

Run my total RNA sample (looking for miRNA) with a DNA ladder on a denaturing gel.
The reason I want to use a DNA ladder is that I can label it with dye and image it and my labeled probe in the Odyssey (thus avoiding the isotope issue)

My question are:
- Will the denaturing gel denature the dsDNA ladder, or do I have to heat it, then put on ice prior to adding to the gel?
- Will the DNA ladder run at the same rate (in relation to size) as my RNA?

 

thanks

alexsunny



#2 alexsunny123

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Posted 24 December 2020 - 03:28 AM

Hello,

 

I was wondering if someone could tell me if this would work;

Run my total RNA sample (looking for miRNA) with a DNA ladder on a denaturing gel.
The reason I want to use a DNA ladder is that I can label it with dye and image it and my labeled probe in the Odyssey (thus avoiding the isotope issue)
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My question are:
- Will the denaturing gel denature the dsDNA ladder, or do I have to heat it, then put on ice prior to adding to the gel?
- Will the DNA ladder run at the same rate (in relation to size) as my RNA?

 

thanks

alexsunny

 

thanks my issue has been fixed.






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