enzyme shear

[brynmcln]

I am shearing for library prep, with the goal being a distribution centered on 500-700bp. However, I am also do not have immediate access to and shearing equipment or a nebulizer. Has anyone tried restriction enzyme fragmentation with good results? Was it easy to achieve the desired size distribution? If so, what kit(s) would you recommend.

Thanks in advance!!

[nucacidhunter]

Fragmentation devices shear DNA randomly which is important to have relatively even coverage of genome. If you use restriction enzyme for fragmentation, your digested DNA will not be random and in best scenario will result in uneven coverage and no coverage in some regions. You can use dsDNA Fragmentase from NEB which is designed for this purpose or use Nextera kit. Also see this thresd: http://seqanswers.com/forums/showthread.php?t=9238

[JamesH]

We just use one of these:

http://www.ebay.com.au/itm/New-2L-Steel-Ultrasonic-Cleaner-Mechanical-Commercial-Grade-Power-60W-/150840947182?pt=AU_Jewellery_Boxes_Supplies&hash=item231ed231ee&_uhb=1

[lorendarith]

There's also an enzyme from Zymo which does the same thing:

http://www.zymoresearch.de/protein/enzymes/dsdna-shearase-plus