title: ChIP-Seq_Multiple myeloma_
design: design_description
library_construction_protocol: NEXTflex ChIP-Seq from Bioo Scientific, 10 cycle PCR at the end and last purification is performed with magnetic beads.
attributes.tag: EXTRACTION_PROTOCOL_TYPE_OF_SONICATOR
attributes.value: Bioruptor Plus (UCD-300)
attributes.tag: CHIP_ANTIBODY_PROVIDER
attributes.value: Diagenode
attributes.tag: CHIP_PROTOCOL_BEAD_TYPE
attributes.value: magnetic beads A and G
attributes.tag: EXPERIMENT_TYPE
attributes.value: Histone H3K4me1
attributes.tag: CHIP_ANTIBODY_LOT
attributes.value: A1863-001P
attributes.tag: CHIP_PROTOCOL
attributes.value: Transcription factor protocol: 100.000 cells per ChIP (5x) and pool at the end. Overnight incubation of chromatin with antibody and A/G magnetic beads. Wash twice with buffer 1, once with buffer 2 and 3 and twice with buffer 4 at 4 degrees. Elute 20 min with elution buffer at RT. Descrosslink 4 hours at 65 degrees. Purify by column purification.
attributes.tag: CHIP_PROTOCOL_BEAD_AMOUNT
attributes.value: 20 µl
attributes.tag: CHIP_PROTOCOL_CHROMATIN_AMOUNT
attributes.value: 5x 100.000 cells = 5x 6,7 µl
attributes.tag: CHIP_PROTOCOL_ANTIBODY_AMOUNT
attributes.value: 0,2 µg
attributes.tag: EXTRACTION_PROTOCOL
attributes.value: Chromatin was extracted using the histone protocol. Cells are crosslinked 10 min in 1% formaldehyde. Washed twice in PBS and dissolved in shearing buffer (containing 1% SDS), such that the final concentration is 15*10^6 cells/ml.
attributes.tag: CHIP_ANTIBODY_CATALOG
attributes.value: #BP140
attributes.tag: CHIP_ANTIBODY
attributes.value: H3K4me1
attributes.tag: EXTRACTION_PROTOCOL_SONICATION_CYCLES
attributes.value: 3x 10 min; 30 sec ON/ 30 sec OFF