Ampure dna purification kit

We performed whole exome sequencing on a cohort of 332 sporadic and mainly late-onset AD cases and 477 elderly controls. DNA was extracted from blood or brain both for cases and controls using standard protocols. Library preparation for next generation sequencing used DNA (between 1 μg and 3 μg) fragmented in a Covaris E210 (Covaris Inc.). Following fragmentation, DNA was end-repaired by 5’phosphorylation, using the Klenow polymerase. A poly-adenine tail was added to the 3’end of the phosphorylated fragment and ligated to Illumina adapters. After purification using an AMPure DNA Purification kit (Beckman Coulter, Inc), adapter-ligated products were amplified. The DNA library was then hybridized to an exome capture library (NimbleGen SeqCap EZ Exome v2.0, Roche Nimblegen Inc. or TruSeq, Illumina Inc.) and precipitated using streptavidin-coated magnetic beads (Dynal Magnetic Beads, Invitrogen). Exome-enriched libraries were PCR-amplified, and then DNA hybridized to paired-end flow cells using a cBot (Illumina, Inc.) cluster generation system. Samples were sequenced on the Illumina HiSeq™ 2000 using 2x100 paired end reads cycles.

DNA was extracted from blood or brain for cases and brain only for controls using standard protocols. Library preparation for next generation sequencing used DNA (between 1 μg and 3 μg) fragmented in a Covaris E210 (Covaris Inc). DNA was end-repaired by 5'phosphorylation, using the Klenow polymerase. A polyadenine tail was added to the 3'end of the phosphorylated fragment and ligated to Illumina adapters. After purification using an AMPure DNA Purification kit (Beckman Coulter, Inc), adapter-ligated products were amplified. The DNA library was then hybridized to an exome capture library (NimbleGen SeqCap EZ Exome v2.0, Roche Nimblegen Inc or TruSeq, Illumina Inc) and precipitated using streptavidin-coated magnetic beads (Dynal Magnetic Beads, Invitrogen). These exome libraries were polymerase chain reaction amplified and then DNA hybridized to paired-end flow cells using a cBot (Illumina, Inc) cluster generation system. Samples were sequenced on the Illumina HiSeq 2000 using 2 × 100 paired-end reads cycles.