Covaris M220 out-of-box use?

[Squall]

Hi all,

We have recently got a Covaris M220 with the NGS Microtube Starter kit.

I am going to use the settings for 500bp peak using the microTUBE AFA Fiber Screw-Cap as advised here: http://covarisinc.com/wp-content/uploads/pn_010166.pdf.

Has anyone used these settings and tube? If so, do you know an ideal DNA concentration to use these settings at? I realise that I will be doing some range-finding experiments, but if I got an idea of a starting point it would be very helpful.

Many thanks.

[cement_head]

Hi all,

We have recently got a Covaris M220 with the NGS Microtube Starter kit.

I am going to use the settings for 500bp peak using the microTUBE AFA Fiber Screw-Cap as advised here: http://covarisinc.com/wp-content/uploads/pn_010166.pdf.

Has anyone used these settings and tube? If so, do you know an ideal DNA concentration to use these settings at? I realise that I will be doing some range-finding experiments, but if I got an idea of a starting point it would be very helpful.

Many thanks.

The stock protocols work really well for what they are - if you deviate from recommended concentrations (protocols on Covaris Website) then you start to have to experiment.

[Squall]

Thanks cement_head.

On the protocol (the one I found anyway) it states to use DNA from 100ng to 5ug - bit of a wide range. I am assuming that the shearing will be more efficient at the lower concentrations, just not sure where to start as the tubes seem quite expensive to do a wide range-finding experiment. Will probably try the middle somewhere.

Thanks!

[cement_head]

If they say the range then its good for the range. We've only had issues with concentractions and volumes outside of recommendations & then what we done is "empirical" testing.

The tubes ARE expensive, but they focus the energy in an exact manner and it gets you the best results...