t4 dna ligase  (Thermo Fisher)

Journal: bioRxiv

Article Title: Robust identification of extrachromosomal DNA and genetic variants using multiple genetic abnormality sequencing (MGA-Seq)

doi: 10.1101/2022.11.18.517160

Figure Lengend Snippet: Experimental procedure, time, and cost of multiple genetic abnormalities sequencing (MGA-Seq). (A) Flowchart of MGA-Seq. Nuclei were cross-linked with 0.5% formaldehyde and then digested with HindIII. 5’ DNA overhangs of digested chromatin fragments were filled in by DNA polymerase and then proximity ligated by T4 DNA ligase. The proximity ligation products were fragmented and then subjected to high-throughput sequencing library construction. After sequencing, all the reads were used to generate chromatin contact matrix for genome structural variation calling. In the sequencing library, the reads without ligation junction “AAGCTAGCTT” were used for the detection of CNV, SNP, small indels (

Article Snippet: In situ proximity ligation Add 27.5 μl of H2O, 3 μl of ATP (adenosine-triphosphate, 10mM), and 10 μl of T4 DNA ligase (Thermo, EL0011) to the reaction system, and placed the tube on the rotating mixers for 2 h at room temperature with rotation at 20 r.p.m.

Techniques: Sequencing, Ligation, Next-Generation Sequencing