Whole-Genome Sequencing: Automated, Nonindexed Library Preparation.
This protocol describes an automated procedure for constructing a nonindexed Illumina DNA library and relies on the use of a CyBi-SELMA automated pipetting machine, the Covaris E210 shearing instrument, and the epMotion 5075. With this method, genomic DNA fragments are produced by sonication, using high-frequency acoustic energy to shear DNA. Here, double-stranded DNA is fragmented when exposed to the energy of adaptive focused acoustic shearing (AFA). The resulting DNA fragments are ligated to adaptors, amplified by polymerase chain reaction (PCR), and subjected to size selection using magnetic beads. The product is suitable for use as template in whole-genome sequencing.
Elaine
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Mardis
W Richard
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McCombie
D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D011088:DNA Ligases / Q000378:metabolism; D015723:Gene Library; D023281:Genomics; D008967:Molecular Biology / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013010:Sonication
D004247:DNA; D011088:DNA Ligases