Bands in gel electrophoresis

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maegovannen
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Bands in gel electrophoresis

Post by maegovannen » Sun Apr 15, 2012 12:02 am

I'm wondering the cause of the following occurance.

A transcription factor was digested with various restriction enzymes, the restriciton fragments were then cloned into a vector, then transcribed and translated and then the fragments were analysed using polyA gel electrophoresis.

There are 3 separate restriction fragements (bands on the gel), 2 and 3 each produce individual bands, however when they're mixed, the gel analysis reveals 3 bands on the gel, 2 of these have migrated the same difference as the bands in the individual fragements of 2 and 3. Can anyone explain this to me? :?: :?: :?:

Thanks in anticipation. :o

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JackBean
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Re: Bands in gel electrophoresis

Post by JackBean » Mon Apr 16, 2012 9:21 am

maegovannen wrote:A transcription factor was digested with various restriction enzymes...


you probably mean gene for transcription factor?

I´m not sure if I understand it correctly, but I would see it that the two fragments are overlapping and thus they combine during electrophoresis giving third band.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

maegovannen
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Re: Bands in gel electrophoresis

Post by maegovannen » Mon Apr 16, 2012 8:09 pm

yes a gene for a transcription factor sorry.

The 2 of the 3 bands are the same as the the fragments they were made from, the 3rd band is inbetween the size of the original fragments.. if that makes sense.

So would it be correct to say that the 2 fragments that were mixed are overlapping?

Thank you again! :)

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JackBean
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Post by JackBean » Tue Apr 17, 2012 7:29 am

If it was combined, it should be bigger.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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