Scale E20 ?

[soleulloa]

Hi all,

I'm working with gDNA from bacteria and from human, doing mate-paired libraries to sequence using SOLID 4.

I want to sequence these samples using a slide divided in 4 parts:
1/4 for bacterial DNA
3/4 for human DNA

I think I have to use E20 to perform emPCR. But I don't know if it is correct.
I'll have to prepare 3 volumes of E20 for human DNA and 1 volume of E20 for bacterial DNA?
Is this correct?

Please, I need your help!

Best regards,
Soledad

[pmiguel]

Do you have an EZ bead system?

The E20 is for ~125 million beads. That is plenty for a quarter slide.

If so, you run it and count the beads you get by optical density of the beads on a spectrophotometer. If in range and you have enough bead to deposit, you are ready to go. Feeling paranoid, run a quick "WFA" (test) run and see what your satays look like.

If you don't have an EZ bead system, you probably don't want to use the EZ bead reagents.

Download the EZ Bead Emulsifier Getting Started Guide (PN 4441486) from the ABI web site for more information.
--
Phillip