Gel for short DNA


Agarose/polyacrilamide composite gel is also called AgPAGE. In case of resolving DNA fragments use the same instruments as for standard agarose gel electrophoresis. In first place mix all components of PAGE except TEMED, then add warm agarose, mix well then add TEMED. The important moment – which temperature the agarose should have, because high temperature induces PA polymerization. If you are not intended to do gradient gel, i.e. you need just the gel, so I think you can use more hot agarose may be 40 – 50C. If you use low melting agarose you should decrease the temperature to about 32-35C. After addition of TEMED allow the gel to polymerize for about 1 hour at room temperature. May be you need optimize the concentration of the agarose and PA to obtain the better resolution. I think agarose should be at least 3.5% and PA at least 15% and may be more. In some protocols there are additional steps of treatment of the mix gel I think this is to make additional cross-links between agarose and PA

Advertisements

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s