Roche 454 Paired End with Covaris Shearing

[LMcSeq]

Hi all,

I'm working on the paired end process using the Roche/454 system for 8kb frags. I'm using the hydroshear for the initial fragmentation, but intend to use the Covaris for the downstream fragmentation step to eliminate the nebulization process.

Does anyone know if there are any adjustments required to the Covaris shearing program (we use the recommended settings for 500bp)?

Thanks!

[Hamid]

Hello,

Just to make sure I provide you the correct information, is your intention to use a hydroshear to shear down to 8kb, and then use the Covaris instrument to shear down to 500bp?

Thank you

hamid

[LMcSeq]

That is correct.

[Hamid]

Hi LMcSeq,

Although you can use the 500bp setting suggested in our protocol (http://covarisinc.com/pdf/pn_400056.pdf), the settings in the attached file works quite well with higher concentrations of DNA up to 50ug in 120ul total sample volume. The setting generate an average size range of 425-475bp, and should be better suited with starting material length of less than 10kb.

Thank you

hamid