Hi-C libraries were generated from 10 million cells by following the in situ Hi-C protocol as mentioned in Chathoth and Zabet (2019) (link). Briefly, crosslinked cells were lysed, and the genome was digested using DpnII (NEB) overnight. The overhangs were filled with Biotin-16-dATP (Jena Bioscience) followed by ligation and de-crosslinking with Proteinase K digestion. The sample was further sonicated using Bioruptor. Biotinylated DNA was pulled down using Dynabeads MyOne Streptavidin T1 beads (Invitrogen 65602). Selected biotinylated DNA fragments ranging from 200 to 500 bp were then ligated with Illumina adaptors (NEB). The libraries obtained from biological replicates were multiplexed and further sequenced at the Oxford Genomics Centre and Edinburgh Genomics (Genepool) using HiSeq 4000.
Biotin 16 datp
Manufactured by Jena Biosciences
Biotin-16-dATP is a nucleotide analog used in molecular biology applications. It contains a biotin molecule attached to the C-16 position of the deoxyadenosine triphosphate. This modification allows for the detection and labeling of nucleic acids during various procedures.
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Lab products found in correlation
3 protocols using biotin 16 datp
1
Hi-C Library Generation from Cell Populations
2
Hi-C Library Generation and Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Hi-C libraries were generated from 10 million cells by following the in situ Hi-C protocol as mentioned by Rao et al. (2014) (link), with minor modifications. Crosslinked cells were lysed, and the genome was digested using DpnII (NEB) overnight. The overhangs were filled with Biotin-16-dATP (Jena Bioscience) followed by ligation and decrosslinking with Proteinase K digestion. The sample was further sonicated using Bioruptor. Biotinylated DNA was pulled down using Dynabeads MyOne streptavidin T1 beads (Life Technologies 65602). Selected biotinylated DNA fragments ranging from 200–500 bp were then ligated with Illumina adaptors (NEB). The libraries obtained from biological replicates were multiplexed and further sequenced at Edinburgh Genomics (Genepool) and Fasteris using HiSeq 4000.
3
Hi-C Library Generation and Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hi-C libraries were generated from 10 million cells by following the in situ Hi-C protocol as mentioned in 26 . Briefly, crosslinked cells were lysed and genome was digested using DpnII (NEB) overnight. The overhangs were filled with Biotin-16-dATP (Jena Bioscience) followed by ligation and de-crosslinking with proteinase K digestion. The sample was further sonicated using Bioruptor. Biotinylated DNA was pulled down using Dynabeads MyOne Streptavidin T1 beads (Life technologies, 65602). Selected biotinylated DNA fragments ranging from 200-500bp were then ligated with illumina adaptors (NEB). The libraries obtained from biological replicates were multiplexed and further sequenced at Oxford Genomics Centre and Edinburgh Genomics (Genepool) using HiSeq4000.
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