GSM1615101: Lib1_Gcn4; synthetic construct; OTHER
The material for sequencing was obtained with a newly developed protocol, BunDLE-seq that enables quantitative measurements of TF (or histones) binding to a pool of thousands of designed sequences, in a single experiment. In this assay, we design a library of sequence variants, up to 200 bp in length, that is then synthesized on Agilent programmable microarrays. Next, we incubate the obtained DNA with a buffer alone (no protein present) or with different concentrations of the examined TF, run the products of this incubation on gel, and extract the DNA from each of the bands detected, corresponding to either naked DNA or DNA bound to different numbers of TF molecules. We amplify the DNA with a unique barcode, marking the originating band, join all samples together and send them to high-throughput sequencing. 10 ng were used for library preparation for sequencing (protocol adopted from(Blecher-Gonen et al. 2013)). The DNA amplified using 14 amplification cycles and sequenced on a 50 bp single read flowcell on Illumina HiSeq-2000 sequencer
GEO Accession
GSM1615101