Illumina suggests hydroshear or nebulizer for the first shearing in the mate-pair library protocol. I was wondering if anyone has used a covaris for this first step or if the covaris can shear to this bp length of 2-5kb. any examples or suggestions would be greatly appreciated.
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Hi Chaos,
Covaris actually has protocols for the 2, 3kb, and 4kb shearing. You can find the protocols and the shearing data at the link below:
2kb- http://covarisinc.com/pdf/pn_400070.pdf
3kb- http://covarisinc.com/pdf/pn_400069.pdf
5kb- http://covarisinc.com/pdf/pn_400071.pdf
Thank you
Hamid -
that is perfect! thank you for the help!Comment
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Originally posted by Hamid View Post
Covaris consumable shears DNA to 6kbp - 20kbp fragments
g-TUBE, the latest innovation from Covaris, is a single-use consumable that enables scientists to shear genomic DNA into selected fragments sizes ranging from 6kbp to 20kbp. The only equipment needed is a compatible bench-top centrifuge and a pipette.
g-TUBE uses centrifugal force (the "g" in g-TUBE) to push the sample through a precisely manufactured orifice in the embedded ruby. This produces shearing forces in the sample that fragment the DNA. Fragment size is selected by adjusting the centrifuge rotor speed which alters the flow rate through the ruby and thus the shearing forces. Higher centrifugation RPM will produce shorter fragments and the protocol provide by Covaris contains the settings needed to select a fragment size between 6kbp and 20kbp.
Remarkably, the entire process takes only 3 minutes and up to 24 samples can be processed simultaneously.Comment
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