Home Abstract Introduction
Cross-linking Cell Lysis and DNA Fragmentation Chromatin immunoprecipitation and elution of DNA cross-link reversal and DNA Purification preparation of DNA for Sequencing Next Generation Sequencing process
KLF1 peaks Distance of promoter peaks Analysis of KLF1 binding to gene body of ZBTB7A

Website for translation of

Laura J. Norton, Alister P.W. Funnell (2017)
KLF1 directly activates expression of the novel fetal globin repressor ZBTB7A/LRF in erythroid cells
Blood Advances 1:685-692

(Translated by Wassal Alhammad)

Experiment: cross-link reversal and DNA Purification

Fourth step is to free up the crosslinked DNA. This step is called cross-link reversal and can be achieved by prolonged heat. The increased temperature will slowly allow formaldehyde to be released from DNA and Protein and therefore results in free DNA. The products are free DNA and KLF1 proteins.

After cross-link reversal, free DNA can be purified using conventional ethanol precipitation method.

Purified DNA at this step is ready for sequencing procedure.

Fig. 7: Construction of cross-link reversal and DNA Purification. increasing heat , on KLF1-bound to DNA, will get KLF1 unbounded from DNA. then, DNA purified by ethanol precipitation method.