Acetoin (98%) and 2,3-butanediol (98%) were purchased from Shanghai Jingchun Reagent (China). D-2,3-butanediol (>96%) was purchased from Tokyo Chemical Industry (Tokyo, Japan), meso-2,3-butanediol (99%) was purchased from Sigma-Aldrich (Sigma, St. Louis, MO, USA). All other chemicals were of analytical grade supplied by Sinopharm Chemical Reagent (Shanghai, China). T4 DNA ligase and DNA marker were purchased from Takara Bio (Dalian, China). TransStart FastPfu DNA Polymerase was purchased from TransGen Biotech (Beijing, China). Plasmid Miniprep Kit was obtained from Zoman Biotech (Beijing, China). Nucleotide sequences were determined by Beijing Genomics institution (Beijing, China).
Dna marker
Manufactured by Takara Bio
Sourced in China, Japan
DNA marker is a molecular tool used to determine the size of DNA fragments in gel electrophoresis. It is a mixture of DNA fragments of known sizes, which can be used as a reference to estimate the size of unknown DNA samples.
Automatically generated - may contain errors
Lab products found in correlation
T4 dna ligase, by Takara Bio (26 mentions)
Restriction enzyme, by Takara Bio (8 mentions)
Dntps, by Takara Bio (6 mentions)
Restriction endonuclease, by Takara Bio (6 mentions)
Dna gel extraction kit, by Takara Bio (5 mentions)
Trizol reagent, by Takara Bio (4 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
T4 dna ligase, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Trizol reagent, by Thermo Fisher Scientific (4 mentions)
Protein marker, by Thermo Fisher Scientific (3 mentions)
Ex taq, by Takara Bio (3 mentions)
Taq dna polymerase, by Takara Bio (3 mentions)
Gel extraction kit, by Takara Bio (2 mentions)
Loading buffer, by Takara Bio (2 mentions)
Dntp mixture, by Takara Bio (2 mentions)
La taq dna polymerase, by Takara Bio (2 mentions)
Sybr green realtime pcr master mix, by Toyobo (2 mentions)
Dual luciferase reporter assay system, by Promega (2 mentions)
Revertra ace qpcr rt kit, by Toyobo (2 mentions)
57 protocols using dna marker
1
Production of 2,3-Butanediol Isomers
2
Recombinant Protein Expression in E. coli
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The strains and vectors used in this study are shown in Table S1 . E. coli Top10 stored in our laboratory was used for all cloning steps. For recombinant proteins expression, pET-21a vector (Novagen, Madison, WI) was used with expression host E. coli BL21(DE3)pLysS (Novagen, Madison, WI). Restriction enzymes, Pyrobest DNA Ploymerase, dNTP, DNA Marker and gel extraction kits were obtained from TaKaRa (Dalian, China). The mouse anti-His monoclonal antibody and enhanced horseradish peroxidase (HRP)-Diaminobenzidine (DAB) chromogenic substrate kit were obtained from Tiangen (Beijing, China). FITC labeled donkey anti-mouse IgG, HRP labeled rabbit anti-mouse IgG, lead acetate, IPTG, and antibiotics were all purchased from Sangon Biotech (Shanghai, China). Pepsin (1:3000) was obtained from Amresco (OH, USA). Tryptone and yeast extract were obtained from Oxoid (Basingstoke, UK). E. coli was cultured in Luria-Bertani (LB) broth (1% tryptone, 0.5% yeast extract and 1% NaCl) supplemented with antimicrobial agents, as necessary.
3
Rat Nasal Allergy Assay Protocol
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Grade II and V OVA were purchased from Sigma (MO, USA). Al(OH)3, IgE, IL-2, and IL-4 ELISA kits were acquired from Elisa biotech (Shanghai, China). Bacterial genome extraction kit and DNA marker were obtained from Takara (Dalian, China). FastPfu PCR mix was procured from Transgene Biotech (Beijing, China). Tris, agarose, EDTA2, H2O, and PCR primers were obtained from Sangon Biotech (Shanghai, China). DNA gel purification kit and AxyPrep DNA gel recovery kit were purchased from Axygen Biosciences (CA, USA). Four kinds of fluorescent dNTPs were acquired from Shanghai Meiji Biotech (Shanghai, China). QuantiFluor-ST blue fluorescence quantitative system was procured from Promega (Wisconsin, USA). Rat nose poisoning apparatus was obtained from Hope Co., Ltd., (Tianjin, China).
4
SSR Marker Development and Diversity Analysis in Rhododendron
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Genomic DNA was extracted from fresh leaves with a modified CTAB (cetyltrimethyl ammonium bromide) method and diluted to 50 ng/μL (Wang et al. 2017 ). A set of 45 R. simsii unigenes containing (AG/CT)n motifs (n ≥ 20) and enough flanking sequences (more than 100 bp) were randomly chosen for prime pair design with online software Primer 3 (Wang et al. 2017 ). In total, twenty SSR markers were developed for analysis of population diversity (Supplemental Table 1 ). The 10 μL SSR-PCR reaction system consisted of 0.2 μM for each primer, 50 ng genomic DNA, and 5 μL 2 × Taq Plus PCR MasterMix (TianGen, Beijing, China). PCR amplification conditions included initial denaturation at 95°C for 10 min, 35 amplification cycles (94°C for 30 s, annealing at optimal temperature for 40 s, and 72°C for 50 s), and a 10 min elongation step at 72°C. The PCR products were separated on 6% (w/v) denaturing poly-acrylamide sequencing gels along with DNA marker (20–500 bp, Takara), and the gels were further visualized by silver staining. PCR products were scored manually, and the genetic parameters were calculated with the ARLEQUIN 3.01 software, including number of alleles (Na) per locus, expected heterozygosity (HE), observed heterozygosity (HO), and deviation from Hardy-Weinberg equilibrium (HWE) (Excoffier et al. 2005 ).
5
Mycotoxin Detection Protocol Development
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The standard mycotoxin powders, the surfactants Tween-20, and the enzyme stabilizer bovine serum albumin (BSA) were obtained from Sigma-Aldrich (St. Louis, MO, United States). Escherichia coli ER2738 competent cells were purchased from Lucigen Corp. (Middleton, WI, United States). The Universal Probes Supermix was supplied by Bio-Rad (Hercules, CA, United States). DNA polymerase (iTaq), Mg2+, dNTPs, 6× loading buffer, and DNA marker were bought from Takara Bio (Beijing, China). All the other reagents used were of analytical grade or better.
The anti-aflatoxins monoclonal antibody 1C11 (mAb 1C11) and V2–5 phage displaying nanobody specific for 1C11 were produced by our team (Zhang et al., 2009 (link); Wang et al., 2013b (link)). A. flavus strain 3.4408 producing a high level of aflatoxins B1 and B2 was used as a standard strain.
The anti-aflatoxins monoclonal antibody 1C11 (mAb 1C11) and V2–5 phage displaying nanobody specific for 1C11 were produced by our team (Zhang et al., 2009 (link); Wang et al., 2013b (link)). A. flavus strain 3.4408 producing a high level of aflatoxins B1 and B2 was used as a standard strain.
6
Comprehensive Biological Sample Processing
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The DNA extraction kit and protease K were purchased from Tiangen Biochemistry Technology Co., Ltd., and the crystal violet and the phosphate buffer solution (PBS) buffer were purchased from Shanghai Shenggong Bioengineering Co., Ltd. The PCR polymerase, restriction endonuclease, and DNA marker were purchased from TaKaRa Company of Japan, and the MALDI-TOFMS mass spectrometer was purchased from Bruker Daltonik Company (Germany). The PCR instrument was purchased from the Applied Biosystems Company of the United States, and the enzyme labeling instrument was purchased from the Biotek Company of the United States. The pulsed gel electrophoresis system was purchased from Bio-Rad Company, the Ultra Micro Spectrophotometer (NanoDrop2000) was purchased from Thermo Company, and the Ultraviolet Gel Imager was purchased from Shanghai Tianneng Company. The −80°C ultra-low temperature refrigerator and incubator were purchased from Thermo Company, while the micro-pipette and centrifuge were purchased from Eppendorf Company (Germany).
7
miRNA Target Expression Assay Protocol
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TRIzol reagent, pMD18-T vector, LA Taq DNA polymerase, dNTP mixture, DNA marker, T4 DNA ligase, XhoI and XbaI were purchased from Takara Bio Inc. (Otsu, Japan). The ReverTra Ace qPCR RT kit and SYBR® Green Realtime PCR master mix (QPK-201) were purchased from Toyobo Co., Ltd. (Osaka, Japan) for reverse transcription quantitative polymerase chain reaction (RT-qPCR). The endotoxin-free plasmid extraction kit was purchased from Tiangen Biotech Co., Ltd. (Beijing, China). The pmirGLO Dual-Luciferase miRNA Target Expression vector and Dual-Luciferase® Reporter Assay system were purchased from Promega Corporation (Madison, WI, USA). Dulbecco's modified Eagle's medium: Nutrient mixture F-12 (DMEM/F12) was purchased from Gibco-BRL (Invitrogen Life Technologies, Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from Hyclone (GE Healthcare, Logan, UT, USA). miR-26b and the negative controls were synthesized by Shanghai Jima International Trading Co., Ltd. (Shanghai, China). The X-tremeGENE small interfering (si)RNA transfection reagent was purchased from Roche Diagnostics (Basel, Switzerland).
8
Isothermal DNA Amplification with Ligation
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The DNA oligonucleotides were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China) and purified by high-performance liquid chromatography. Taq DNA ligase (40 U μL−1), 10 × Taq DNA ligase reaction buffer (200 mM Tris-HCl, 250 mM KAc, 100 mM Mg(Ac)2, 100 mM DTT, 10 mM NAD, 1% Triton X-100, pH 7.6), Bst 2.0 DNA polymerase (8 U μL−1) and 10 × ThermoPol reaction buffer (200 mM Tris-HCl, 100 mM (NH4)2SO4, 500 mM KCl, 20 mM MgSO4, 1% Tween-20, pH 8.8) were all purchased from New England Biolabs (Beijing, China). SYBR Green I was obtained from Generay Biotech. Co., Ltd. (Shanghai, China). DNA marker, 6 × loading buffer and dNTPs were purchased from Takara (Dalian, China). TIANamp Genomic DNA Kit was purchased from Tiangen Biotech Co., Ltd. (Beijing, China). The nuclease-free water was purchased from Thermo Fisher Scientific Inc (Vilnius, Lithuania) and used in all ligation reaction and ligation-initiated LAMP assays. All the reagents were of analytical grade and used without further purification. All solutions were prepared with ultrapure water from Millipore Milli-Q water purification system (Millipore, USA). The detailed oligonucleotide sequences were listed in Table S1 (Supporting Information).
9
DNA Oligonucleotide Synthesis and Cell Line Experiments
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All DNA oligonucleotides (Supporting Information, Table S1 ) were synthesized and purified by Sangon (Shanghai, China). The DNA oligonucleotides were dissolved in 1 × Tank buffer (20 mM Tris,125 mM NaCl, 20 mM KCl, pH 7.5) and diluted in appropriate buffer prior to use or stored at −20 °C. DNA Marker, GoldView, SYBR Premix Ex TaqTM II and PrimescriptTM RT reagent Kit with gDNA Eraser were purchased from TaKaRa (Dalian, China). Dulbecco’s Modified Eagle Medium (DMEM) and Fetal Bovine Serum (FBS) were obtained from Gibco (Shanghai, China). CCK-8 assay was obtained from Dojindo (Shanghai, China). Phospho-HER2/ErbB2 Antibody Sampler Kit and β-Actin polyclonal antibody were obtained from Cell Signaling Technology (Shanghai, China). HRP anti-mouse and rabbit IgG were from Beyotime (Jiangsu, China).New Super ECL Assay was from KeyGen (Nanjing, China). RIPA (50 mM Tris (pH 7.4), 150 mM NaCl, 1% Triton x-100, 1% sodium deoxycholate, 0.1% SDS, sodium orthovanadate, sodium fluoride, EDTA, and leupeptin), phenylmethanesulfonyl fluoride (PMSF) and Bradford protein dye reagent were used in western blot. SK-BR-3and MDA-MB-231 (the human breast cancer cell lines) cells were obtained from the American Type Culture Collection (ATCC) (Rockville, MD, USA). All other reagents were of analytical grade, and Millipore-Q water (≥18 МΩ) was used in all experiments.
10
Construction of Rel Gene Deletion Mutant
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S. diastatochromogenes 1628 was isolated and stored in the China General Microbiological Culture Collection Center (CGMCC no. 2060). E. coli JM109 was used to construct the plasmids. The pKC1139 was used to delete the Rel gene. The primers were listed in Table S1 in Supplemental File 1. All Escherichia coli strains were grown in the Luria Bertani (LB) and 2×yeast extract tryptone (2×YT) medium at 37°C supplemented with antibiotics if necessary. S. diastatochromogenes 1628 was cultured in the GYM medium and appropriate antibiotics were added at 28°C on a rotary shaker at 180 rpm. The methylation-deficient strain of E. coli , ET12567/pUZ8002, was used as the donor for plasmid transfer by intergeneric conjugation to Streptomyces. Conjugation and selection of exconjugants were performed on the solid mannitol soya flour (MS) agar medium for sporulation. The LA-Taq DNA polymerases, T4 ligase, DNA marker, and other related enzymes were purchased from TaKaRa, Kusatsu, Japan. The genome of S. diastatochromogenes , the plasmids extraction, and purification of DNA fragments was performed by relative kits according to the kit proposal from Kakara (Dalian, China).
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