Help an undergrad with the basics?

[undergrad]

It would be great if somebody had the patience to explain the 454 Roche preparation procedure out to me.

My understanding is that I have to amplify my DNA samples using site specific primers (in my case 28s LSU). These primers are modified to contain the 454 A and B primers and one of the Roche barcode adaptors. Following PCR is purification and emulsion, then shipping to a 454 processing lab.

Also, if there's a paper describing this, that would be great! The papers I've read have been vague and my lab members have not done this before to help me. Thanks!

[GraemeFox]

Try the Roche documents:

'System Research Applications Guide' (there are FLX & Junior versions) and the '454 Sequencing System Guidelines for Amplicon Experimental Design'.

Both should be Google-able and give you a good idea of how the sequencing process works.

Hope that helps