[XIAOXIAO]

thanks! i will try 1:1

[liuqy]

Thanks much for the tips. I was always bothered by the primer dimer when used NEB kits for library prep. The AMPure XP worked fabulously for size selection, however didn't work for getting rid of the primer dimer. I had to use Pippin Prep to clean-up the products, which caused more than half loss. After switched to Nextera kit, no more primer dimer problem, but primer contamination presented sometimes. The manufacturer's suggestion is only 25uL beads for 50uL products. I would try your 1.8:1 ratio next time.

You may try NVIGEN MagVigen. It's observed that AmPure XP lags MagVigen in primer dimer/adapter dimer cleanup. You can check these different contamination levels (>200%, >100%, >50%, 20%) (https://www.nvigen.com/dna-clean-up/)

Disclosure: I work at NVIGEN. We cordially invite you to try MagVigen. In case you find problems using it, we will be happily work with you to troubleshoot or fine tune it to meet your needs.

[liuqy]

Hey if you used AmPure beads for purification, what is the ratio of library versus beads for purificaiton?

There is a charter picture here, which demonstrates beads volume vs sample volume (note: not sample to beads). NVIGEN MagVigen uses the same protocols as AmPure XP.

https://www.nvigen.com/dna-size-selection-tutorial/#ratio