DNA affinity purification sequencing (DAP-seq) couples in vitro expression of transcription factor (TF) and binding to a genomic DNA library with next-generation sequencing, providing a robust method to identify genome-wide transcription factor binding sites (TFBS) on endogenous genomic DNA. Here we...
coli, the desired TCTP shRNA-resistant gene was verified by DNA sequencing. Affinity purification of fusion protein expressed from E. coli pET-41a (+) vector expressing GST-Rad51 or pBad/His A vector expressing 6xHis-TCTP (Invitrogen) was transformed into BL21 E. coli, the transformed ...
The protein sequences contained an N-terminal poly- histidine-tag which was always removed after purification through an inserted PreScission Protease recognition cleavage site. All sequences were checked by sequencing analysis. The protein variants were checked and corrob- orated by Sanger sequencing ...
A Simple Method for Determining Compound Affinity and Chemical Yield From DNA-encoded Library SelectionsRelease Time:2020-06-18 Abstract: DNA-encoded libraries (DELs) can contain billions of unique chemical species; selecting against such large inputs, it is typical to find more candidate binders th...
Our sequencing studies revealed a unique mutation (C1462A) in Stat5b of NOD. The mutation is within the DNA binding domain of the Stat5b protein, and the residue is a conserved amino acid in all the seven members of the STAT gene family as well as in various animal species including mic...
(Qiagen, Germany) according to the manufacturer’s protocol, including the optional on-column DNAse treatment. After elution, the RNA was treated with Turbo DNAse (Applied Biosystems, USA) according to the manufacturer’s manual followed by a second round ofpurificationusing the RNeasy Mini Kit....
DNA affinity purification sequencing (DAP-seq) has been shown to contribute to ways of making pure some of the protein with rare sequence-specific DNA binding and can help understand the molecular dynamics in barley under varying exposure time to drought. The present study aimed at identifying ...
Chou et al., 1996, “Affinity methods for purification of DNA sequencing reaction products for mass spectrometric analysis,” Rapid Commun. Mass. Spect. 10:1410-1414.;:1410-1414.Chou et al., Affinity methods for purification of DNA sequencing reaction products for mass spectrometric analysis....
After prioritizing the top 22 TFs for further validation, we identified DNA‐binding sites and predicted target genes for 16 out of the 22 TFs tested using DNA affinity purification followed by sequencing (DAP‐seq). Enrichment analyses of targets supported the co‐expression prediction for most ...
For the peptide sequencing of FMBP-1, the gel was stained with CBB, and the FMBP-1 band was cut out and treated for sequencing. EMSA EMSAs were carried out as described by Takiya et al. (21). The ol...Inomata Y,Kawaguchi H,Hiramoto M,Wada T Handa H Anal. Direct purification of ...