E0554

Manufactured by New England Biolabs

Sourced in Hong Kong

The E0554 is a DNA ligase enzyme from T4 bacteriophage. It catalyzes the formation of phosphodiester bonds between adjacent 3'-hydroxyl and 5'-phosphate termini in double-stranded DNA molecules.

Automatically generated - may contain errors

Lab products found in correlation

Sanger sequencing, by New England Biolabs (1 mentions) Pcmv6 ac bap1 plasmid, by OriGene (1 mentions)

3 protocols using e0554

Mutagenesis of LSD1 Protein K661A

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Mutagenesis was performed according to manufacturer’s protocol (NEB, E0554) using HA-LSD1 plasmid as template, and confirmed via Sanger sequencing by Eurofins Scientific. LSD1 K661A substitution primers were generated using NEBaseChanger.
Primer sequences:
forward, CAACCTTAACgcGGTGGTGTTGTG
reverse, CCAAATCCCATCCTTTGG
Annealing temperature: 58°C

Miller S.A., Policastro R.A., Savant S.S., Sriramkumar S., Ding N., Lu X., Mohammad H.P., Cao S., Kalin J.H., Cole P.A., Zentner G.E, & O’Hagan H.M. (2019). Lysine-specific demethylase 1 mediates AKT activity and promotes epithelial-mesenchymal transition in PIK3CA mutant colorectal cancer. Molecular cancer research : MCR, 18(2), 264-277.

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Lentiviral Expression of BAP1 Variants

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Full-length BAP1 complementary DNA was amplified by PCR from pCMV6-AC BAP1 plasmid (SC117256; Origene) and cloned into the lentiviral plasmid pCCL-CMV-flT vector. Vectors expressing BAP1-mutant constructs were generated by site-directed mutagenesis (E0554; New England Biolabs) of the pCCL-CMV-BAP1 vector as previously described (23 ).

Ishii Y., Kolluri K.K., Pennycuick A., Zhang X., Nigro E., Alrifai D., Borg E., Falzon M., Shah K., Kumar N, & Janes S.M. (2021). BAP1 and YY1 regulate expression of death receptors in malignant pleural mesothelioma. The Journal of Biological Chemistry, 297(5), 101223.

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Cloning and Mutagenesis of Human WRN and Zebrafish wrn

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Human WRN cDNA was synthesized and cloned into the pCDH-EF1 vector (SBI, CD550A-1) between EcoRI and NotI restriction sites. WRN mutagenesis followed the Q5 site-directed mutagenesis instructions (NEB, E0554) Different WRN mutagenesis PCR primers were designed using the NEB online design software NEBase Changer™ (http://nebasechanger.neb.com/) and can be found in Supplementary table 1.
Zebrafish wrn cDNA was synthesized and cloned into the Z-pBluescript II vector (provided by Prof. Zhao Hui, School of Biomedical Sciences, the Chinese University of Hong Kong, Hong Kong) between EcoRI and BamHI restriction sites. Primers were synthesized in BGI Genomics (Beijing, China) and can be found in Supplementary table 1.

Tian Y., Wang W., Lautrup S., Zhao H., Li X., Law P.W., Dinh N.D., Fang E.F., Cheung H.H, & Chan W.Y. (2022). WRN promotes bone development and growth by unwinding SHOX-G-quadruplexes via its helicase activity in Werner Syndrome. Nature Communications, 13, 5456.

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