title: GSM2109264: shATXN7L3_H2Bub; Homo sapiens; ChIP-Seq
library_construction_protocol: Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with antibody. Antibody-histone-DNA complexes were harvested using protein G magnetic dynabeads (ThermoFisher Scientific, Ref-10004D, Lot-163538810) CHIP libraries were prepared using a modified Kapa LTP Library Preparation kit (KAPA Biosystems, Wilmington, MA) protocol for Illumina Platforms. 10ng of sheared DNA was used to repair the ends of the damaged fragments using a proprietary master mix. The resulted blunted fragments had to be 3’ A-Tailing using a proprietary mixture of enzymes to be able to ligate to the specific NexTflex adaptors from Bioo Scientific (Bioo Scientific, Austin, TX). For each of the steps (End Repair, 3’A Tailing, and Adaptor Ligation was followed by column clean up (Qiagen, Valencia, CA). After adapter ligation, DNA enrichment was performed using Kapa HiFi Hot Start Ready mix PCR, and a cocktail of primer mix. (1 cycle at 98°C for 45 seconds; 4 cycles of 98°C for 15 seconds, 60°C for 30 seconds, and 72°C for 30 seconds; 1 cycle at 72°C for 1 minute), and purified with AmpureXP beads (Beckman Coulter, Indianapolis. IN).The final library validation was checked using the 2200 TapeStation from Agilent Technologies (Agilent, Santa Clara, CA). Libraries were quantified using a Kapa Library Quantification Kit (KAPA Biosystems) and loaded in a flow cell for cluster generation using the Illumina cBOT (Illumina, San Diego, CA) at final concentration of 10pM
attributes.tag: GEO Accession
attributes.value: GSM2109264