T4 dna ligase

Manufactured by Beyotime

Sourced in China

T4 DNA ligase is an enzyme used in molecular biology to join DNA fragments together. It catalyzes the formation of phosphodiester bonds between adjacent 3'-hydroxyl and 5'-phosphate ends in double-stranded DNA, DNA-RNA hybrids, or single-stranded DNA. The enzyme is derived from the T4 bacteriophage.

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Lab products found in correlation

Dna polymerase for pcr, by Beyotime (2 mentions) Nickel nitrilotriacetic acid ni nta superflow column, by Transgene (2 mentions) Pet 28 a expression plasmid, by Sangon (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Nanodrop 2000, by Thermo Fisher Scientific (2 mentions) Phi29 dna polymerase, by Beyotime (1 mentions) Hydrogen peroxide h2o2, by Beyotime (1 mentions) Milli q 18 mω, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Sangon (1 mentions) Dntps, by Sangon (1 mentions) Transit mrna, by Mirus Bio (1 mentions) Megascript t7 kit, by Thermo Fisher Scientific (1 mentions) Pcdna 3.4 topo ta cloning kit, by Thermo Fisher Scientific (1 mentions) Ambion megaclear spin columns, by Thermo Fisher Scientific (1 mentions) Antarctic phosphatase, by New England Biolabs (1 mentions) Ni nta agarose, by Sangon (1 mentions) Bamhi, by Beyotime (1 mentions) Taq dna polymerase, by Beyotime (1 mentions) Hyaluronic acid sodium salt from streptococcus equi, by Merck Group (1 mentions) Oligonucleotide primers, by Sangon (1 mentions)

8 protocols using t4 dna ligase

DNA Purification and Enzymatic Assay

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The DNA samples used in this study were obtained from Sangon Biotech (Shanghai, China) for experimental purposes; the DNA sequences and modifications are listed in Table 1. All DNA samples were purified by HPLC, stocked in 1× TE buffer (pH = 7.8), and stored at −20 °C.
T4 DNA ligase and phi29 DNA polymerase were purchased from Beyotime Biotech (Shanghai, China). Beyotime Biotech also provided a 10× T4 DNA ligation buffer and 10× phi29 DNA polymerase reaction buffer. Streptavidin immunomagnetic beads (IMBs, 10 mg/mL, 0.5 μm), dNTPs, and BSA were purchased from Sangon Biotech (Shanghai, China). Hemin, 3, 3′, 5, 5′-tetramethylbenzidine (TMB), and hydrogen peroxide (H₂O₂) were purchased from Beyotime Biotech. All other chemical reagents were bought from Sangon Biotech and were of analytical grade. All solutions in this study were prepared using ultrapure water sourced from Milli-Q (18 MΩ, Merck Millipore, Shanghai, China). The buffers used in this method are listed as follows:
Binding Buffer (10×): 100 mM Tris-HCl, 500 mM MgCl₂, 500 mM NaCl.
Enzyme Activity Buffer: 100 mM Tris Base, 120 mM NaCl, 10 mM MgCl₂, 100 mM KCl.
Substrate Buffer: 200 mM Na₂HPO₄, 100 mM citric acid.

Wang Y., Xiao J., Lin X., Waheed A., Ravikumar A., Zhang Z., Zou Y, & Chen C. (2023). A Self-Assembled G-Quadruplex/Hemin DNAzyme-Driven DNA Walker Strategy for Sensitive and Rapid Detection of Lead Ions Based on Rolling Circle Amplification. Biosensors, 13(8), 761.

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Cloning and Expression of Human TRAIL in ADSCs

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Full-length human TRAIL cDNA was cloned into the pcDNA3.3-TOPO plasmid using a pcDNA 3.4 TOPO TA Cloning Kit (#A14697; Thermo Fisher Scientific, Inc.) as shown in Fig. 1A. In brief, TRAIL cDNA was amplified from MIGR1-TRAIL-GFP as described by Wiley et al (18 (link)) and then sub-cloned into the pcDNA3.3-TOPO plasmid. The TRAIL cDNA was connected to the pcDNA 3.3-TOPO plasmid with T4 DNA ligase (#D7006; Beyotime Institute of Biotechnology, Beijing, China). After colony polymerase chain reaction (PCR) amplification (the template was bacterium suspension) and DNA sequencing confirmation (performed by Sangon Biotech Co., Ltd., Wuhan, China), this plasmid containing TRAIL cDNA was used for tail PCR and modified with the MEGAscript T7 kit (Ambion; Thermo Fisher Scientific, Inc.). Then, modified TRAIL mRNA was isolated with Ambion Anti-Reverse Cap Analog (ARCA; #AM8045) and purified with Ambion MEGAclear spin columns (Thermo Fisher Scientific, Inc.) and treated with Antarctic Phosphatase (New England Biolabs, Ipswich, MA, USA) to remove residual 5′-triphosphates. The transfection of the TRAIL plasmid into ADSCs was conducted using TransIT-mRNA (Mirus Bio LLC., Madison, WI, USA) according to the manufacturer' instructions and ADSCs transfected with TRAIL-cDNA were defined as TRAIL-ADSCs.

JING H.X., DUAN D.J., ZHOU H., HU Q.M, & LEI T.C. (2016). Adipose-derived mesenchymal stem cell-facilitated TRAIL expression in melanoma treatment in vitro. Molecular Medicine Reports, 14(1), 195-201.

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Recombinant Protein Expression in E. coli

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E. coli BL21 (DE3), plasmid vector pET15b, and nickel-nitrilotriacetic acid agarose (Ni-NTA agarose) were purchased from Sangon (Shanghai, China). Spin miniprep kit and gel extraction kit were obtained from Bioman (Fuzhou, China). The restriction enzymes NdeI and BamHI, Taq DNA polymerase, and T4 DNA ligase were from Beyotime (Shanghai, China).

Zhong R., Gao L., Chen Z., Yuan S., Chen X, & Zhao C. (2021). Chemoenzymatic synthesis of fucosylated oligosaccharides using Thermosynechococcus α1–2-fucosyltransferase and their application in the regulation of intestinal microbiota. Food Chemistry: X, 12, 100152.

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Bacterial Expression and Purification of Glycosaminoglycans

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DNA extraction kits, DNA polymerase for PCR, and T4 DNA ligase were purchased from Beyotime Biotech Co. Ltd. (Haimen, China). The PET-28(a) expression plasmid, restriction endonucleases, oligonucleotide primers, and competent cells, including E. coli DH5α and BL21 (DE3), were provided from Sangon Bioengineering Co. Ltd. (Shanghai, China). A nickel-nitrilotriacetic acid (Ni-NTA) superflow column was supplied by TransGen Biotech Co. Ltd. (Beijing, China). The DNA ladder and protein molecular weight marker were obtained from Detai Biologics Co. Ltd. (Nanjing, China). CS-A from the bovine trachea, CS-C from shark cartilage, and hyaluronic acid (HA) sodium salt from Streptococcus equi were purchased from Sigma Co. Ltd. (St. Louis, MO, USA). Kanamycin sulfate, IPTG, and all other reagents used in the experiments were of analytical grade and ordered from Aladdin Co. Ltd. (Shanghai, China).

Zhou L.J., Guo L.B., Wei W., Lv Z.X, & Zhang Y.W. (2021). A Novel Chondroitin AC Lyase With Broad Substrate Specificity From Pedobacter rhizosphaerae: Cloning, Expression, and Characterization. Frontiers in Bioengineering and Biotechnology, 9, 808872.

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Protein Expression and Purification Protocol

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T4 DNA ligase and Lipo8000 transfection reagent were purchased from Beyotime Biotechnology (Shanghai, China). 2 × Taq Plus Master Mix (Dye Plus) and T4 polynucleotide kinase was purchased from Vazyme Biotechnology (Nanjing, China). The KOD-Plus Mutagenesis Kit and KOD-Plus Kit were from TOYOBO (Osaka, Japan). Ni²⁺-NTA Superflow resin was purchased from Qiagen Inc. (Hilden, Germany). Dynabeads Protein G, 4,6-diamidino-2-phenylindole (DAPI), MitoTracker, Pierce Silver Stain kit, and Alexa Fluor 488-conjugated secondary antibody were obtained from Thermo Scientific (Waltham, MA, USA). [¹⁴C]Thr, [¹⁴C]Ser and [¹⁴C]Tyr were obtained from Perkin Elmer Inc. (Waltham, MA, USA). PrimeScript RT Master Mix and PrimeSTAR Max DNA Polymerase were obtained from TaKaRa (Kyoto, Japan). Trelief Prestained Protein Ladder and oligonucleotide primers were obtained from Tsingke (Shanghai, China). Competent E. coli BL21(DE3) were purchased from Weidi Biotechnology (Shanghai, China). Serum-free Cell Cryopreservation Medium was obtained from Epizyme Biomedical Technology (Shanghai, China). Anti-c-Myc magnetic beads, kanamycin sulfate and ampicillin sodium were purchased from MedChemExpress (MCE, New Jersey, USA).

Peng G.X., Mao X.L., Cao Y., Yao S.Y., Li Q.R., Chen X., Wang E.D, & Zhou X.L. (2022). RNA granule-clustered mitochondrial aminoacyl-tRNA synthetases form multiple complexes with the potential to fine-tune tRNA aminoacylation. Nucleic Acids Research, 50(22), 12951-12968.

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Plasmid Cloning and Transformation

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Plasmid pUC19, DH5α competent cells, Taq DNA polymerase, X-gal, and dNTPs were purchased from Tiangen Biotec Co., Ltd. (Beijing, China). Vent (exo^-) DNA polymerase and restriction enzymes (HindIII, EcoRI, and PstI) were obtained from New England Biolabs Inc (Ipswich, MA, USA). SYBR® gold nucleic acid gel stain was purchased from Invitrogen, Ltd. (Paisley, UK). T4 DNA ligase and T4 polynucleotide kinase were purchased from Beyotime (Shanghai, China). Plasmid small-extraction kit and 40% acrylamide (acrylamide:bis-acrylamide 19:1) were from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). The E.Z.N.A.® Cycle-Pure Kit was obtained from Omega Biotek Inc (Georgia, USA). All oligonucleotides used in this study were purchased from Suzhou Beixin Biotechnology Co., Ltd. Ultrapure water was prepared using double distilled water (UPW, 18 MΩ·cm).
The following equipments were used: PAGE electrophoresis system (Junyi JY200C, Beijing, China), Micro high-speed centrifuge (Xiangyi TG16-W, Changsha, China), Microvolume UV–Vis spectrophotometer (NanoDrop2000 Thermo Fisher, Boston, Massachusetts, USA), ChemiDoc™ MP Imaging System (BioRad, Hercules, California, USA), MJ MiNi Thermal Cycler (Bio-Rad PTC-1148, Hercules, California, USA), and High-speed refrigerated centrifuge (Xiangyi TGL-16, Changsha, China).

Kan Y., Jin Z., Ke Y., Lin D., Yan L., Wu L, & He Y. (2022). Replicative bypass studies of l-deoxyribonucleosides in Vitro and in E. coli cell. Scientific Reports, 12, 21183.

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Recombinant Protein Expression in E. coli

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Oligonucleotide primers, DNA polymerase for PCR, and T4 DNA ligase were obtained from Beyotime Biotech Co. Ltd (Haimen, China). The PET-28a expression plasmid, DNA extraction kit, and E. coli DH5α and BL21 (DE3) were purchased from Sangon Bioengineering Co. Ltd (Shanghai, China). Restriction endonucleases and nickel-nitrilotriacetic acid (Ni-NTA) superflow column were provided by Transgen Biotech Co. Ltd (Beijing, China). DNA ladder and protein molecular weight marker were obtained from Detai Biologics Co. Ltd (Nanjing, China). Kanamycin sulfate, sodium heparin, and isopropyl-b-D-thiogalactopyranoside (IPTG) were ordered from Aladdin Co. Ltd (Shanghai, China). All other reagents used in the experiments were analytical grade and supplied by Sigma-Aldrich Co. Ltd (St. Louis, MO).

Gao L.W., Zhu H.T., Liu C.Y., Lv Z.X., Fan X.M, & Zhang Y.W. (2020). A highly active heparinase I from Bacteroides cellulosilyticus: Cloning, high level expression, and molecular characterization. PLoS ONE, 15(10), e0240920.

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Synthesis and Characterization of DNA-Functionalized Gold Nanoparticles

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T4 DNA ligase, 10 × T4 DNA buffer and bovine serum albumin were purchased from Beyotime Biotechnology (Shanghai, China). Exonuclease I, exonuclease III, phi29 DNA polymerase and dNTPs were purchased from Novoprotein Scientific Inc. (Suzhou, China). HAuCl₄, sodium citrate, NaCl, sodium dodecyl sulfate (SDS), (2-carboxyethyl) phosphine hydrochloride (TCEP), uranyl acetate and quercetin were purchased from Aladin Ltd (Shanghai, China). PBS, MTT cell proliferation and cytotoxicity detection kit, annexin V-FITC/PI apoptosis detection kit and hematoxylin–eosin (HE) dye solution were purchased from Keygen Biotech (Nanjing, China). Hsp27 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Jin Yibai Biological Technology (Nanjing, China). DNA ladder marker was purchased from Takara Biomedical Technology (Beijing) Co., Ltd. (Beijing, China). All DNA strands were synthesized and purified by Sangon Biotech Co., Ltd (Shanghai, China), and their specific sequences and label information were listed in Additional file 1: Table S1.

Yang Y., Cai X., Shi M., Zhang X., Pan Y., Zhang Y., Ju H, & Cao P. (2023). Biomimetic retractable DNA nanocarrier with sensitive responsivity for efficient drug delivery and enhanced photothermal therapy. Journal of Nanobiotechnology, 21, 46.

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