Methods and apparatuses for nucleic acid shearing by sonication
Methods and kits for preparing nucleic acid fragments from a sample of purified nucleic acid are provided. Alternatively, chromatin or other long polymers can be sheared with similar methods and kits.
1. A method for preparing a gene fragment library, the method comprising:
a) providing a suspension comprising a sample of purified genomic nucleic acid and monodisperse particles, wherein the suspension is an aqueous solution without nonaqueous solvent:
b) sonicating the suspension, thereby generating nucleic acid fragments:
c) collecting the nucleic acid fragments; and
d) preparing a gene fragment library by ligating the nucleic acid fragments to nucleic acid adaptors.
2. The method of claim 1 , wherein the collecting step does not include chemical extraction of the nucleic acid fragments.
3. The method of claim 1 , wherein the sample does not include cell lysates.
4. The method of claim 1 , wherein the particles have a size of about 0.1 micrometers to about 100 micrometers.
5. The method of claim 1 , wherein the suspension contains about 0.1% to about 20% of the particles.
6. The method of claim 1 , wherein the particles comprise at least one of glass, borosilicate glass, silica, soda lime glass, carbamide, metal, metal oxide, ceramic, and metal-coated ceramic.
7. The method of claim 1 , wherein the particles are non-magnetic or paramagnetic.
8. The method of claim 1 , wherein the sonicating step is performed by focusing acoustic sonication.
9. The method of claim 1 , wherein the time for the sonicating step is less than 5 minutes to generate about 60 to about 90 base pair nucleic acid fragments from the purified genomic nucleic acid.
10. The method of claim 1 , further comprising sequencing the nucleic acid fragments.
11. The method of claim 1 , further comprising hybridizing the nucleic acid fragments to target nucleic acids located on a microarray.