All oligonucleotides used in this study were synthesized and purified by Tsingke Biotechnology Co., Ltd. (Wuhan, China) and Sangon Biotech Co., Ltd. (Shanghai, China). Vent (exo-) DNA Polymerase (Vent), Nt.BstNBI nicking enzyme (Nt.BstNBI), 10× NEBuffer 3.1 (1 M NaCl, 500 mM Tris–HCl, 100 mM MgCl2, 1 mg/ml BSA, pH 7.9), 10× ThermoPol Reaction Buffer (200 mM Tris–HCl, 100 mM (NH4)2SO4, 100 mM KCl, 20 mM MgSO4, 0.1% Triton X-100, pH 8.8) were purchased from New England Biolabs Inc. (Beijing, China). Deoxynucleotides (dNTPs, 2.5 mM) was purchased from Tiangen Biotech Co. Ltd. (Beijing, China). Agarose, 5× TBE Buffer, 4S Red Plus Nucleic Acid Stain, 6× DNA Loading Dye and DNA Marker A (25–500 bp) were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). The concentration of DNA oligonucleotides was measured using a NanoDrop 2000 UV–vis spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). The sequences of all oligos are listed in Supplementary Table S1 . The secondary structure of TJDHs at 55°C predicted by NUPACK (http://nupack.org/ ) (29–32 (link)) is shown in Supplementary Figure S5 .
Tbe buffer
Manufactured by Sangon
Sourced in China
5× TBE Buffer is a concentrated solution used for preparing and running agarose gel electrophoresis. It provides the necessary ionic environment to facilitate the separation and migration of nucleic acid molecules during electrophoresis.
Automatically generated - may contain errors
Lab products found in correlation
Nebuffer 3, by New England Biolabs (1 mentions)
Nanodrop 2000 uv vis spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Thermopol reaction buffer, by New England Biolabs (1 mentions)
Dna loading dye, by Sangon (1 mentions)
4s red plus nucleic acid stain, by Sangon (1 mentions)
Agarose, by Sangon (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Rnase inhibitor, by Takara Bio (1 mentions)
Dna probe, by Sangon (1 mentions)
Generuler ultra low range dna ladder, by Thermo Fisher Scientific (1 mentions)
Dntp mixture, by Sangon (1 mentions)
Chamq universal sybr qpcr master mix, by Vazyme (1 mentions)
Pbs buffer, by Sangon (1 mentions)
Taq pcr master mix, by Vazyme (1 mentions)
Rnase free ddh2o, by Sangon (1 mentions)
El tmb chromogenic reagent kit, by Sangon (1 mentions)
Tween 20, by Sangon (1 mentions)
Fast digest mvai, by Thermo Fisher Scientific (1 mentions)
4 protocols using tbe buffer
1
Synthesizing and Characterizing DNA Hairpins
2
Sensitive RNA Detection Assay
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T4 RNA ligase 2, phi29
DNA polymerase, and endonuclease IV (Endo IV) were purchased from
New England Biolabs (Ipswich, MA, USA). HPLC-purified miRNAs, RNase
inhibitor, and RNase-free water were purchased from Takara Biotechnology
Co., Ltd. (Dalian, China). The DNA probes, 5× TBE buffer (225
mM Tris–boric acid, 50 mM EDTA, pH 8.0), and dNTP mixture were
obtained from Sangon Biotech Co., Ltd. (Shanghai, China). GeneRuler
Ultra Low Range DNA Ladder was obtained from Thermo Fisher Scientific
Inc. (Waltham, MA, USA). RPMI 1640 medium, penicillin, 15% heat-inactivated
fetal bovine serum, and streptomycin were obtained from Thermo Scientific
HyClone (MA, USA). Sequences of DNA probes and miRNAs are given in
Table S1,Supporting Information . All other
reagents were of analytical grade and used without additional purification.
RNase-free water was used in all experiments.
DNA polymerase, and endonuclease IV (Endo IV) were purchased from
New England Biolabs (Ipswich, MA, USA). HPLC-purified miRNAs, RNase
inhibitor, and RNase-free water were purchased from Takara Biotechnology
Co., Ltd. (Dalian, China). The DNA probes, 5× TBE buffer (225
mM Tris–boric acid, 50 mM EDTA, pH 8.0), and dNTP mixture were
obtained from Sangon Biotech Co., Ltd. (Shanghai, China). GeneRuler
Ultra Low Range DNA Ladder was obtained from Thermo Fisher Scientific
Inc. (Waltham, MA, USA). RPMI 1640 medium, penicillin, 15% heat-inactivated
fetal bovine serum, and streptomycin were obtained from Thermo Scientific
HyClone (MA, USA). Sequences of DNA probes and miRNAs are given in
Table S1,
reagents were of analytical grade and used without additional purification.
RNase-free water was used in all experiments.
3
ASFV p30 Sandwich ELISA Protocol
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All oligonucleotides (HPLC purified) were synthesized and modified by Sangon Biotechnology Co., Ltd. (Shanghai, China). D-PBS buffer, 20× PBS buffer, 5× TBE buffer, RNase-free ddH2O, Tween-20, Bovine Serum Albumin (BSA), HRP-labeled Streptavidin (SA-HRP), and EL-TMB Chromogenic Reagent kit were purchased from Sangon Biotechnology Co., Ltd. (Shanghai, China). ASFV p30 recombinant protein, p30 monoclonal antibody, porcine reproductive and respiratory syndrome virus (PRRSV) recombinant N protein, staphylococcus aureus enterotoxin type A recombinant protein (SEA), and pig immunoglobulin G (IgG) were purchased from Aiyi Biotechnology. (Nanjing, China). 6× His tag peptides were bought from Synpeptide Biotechnology Co., Ltd. (Nanjing, China). Swine serum was sourced from Solarbio Science & Technology Co., Ltd. (Beijing, China). NHS (N-hydroxysuccinimide)-Activated Magnetic Beads (NHS-MB) were purchased from Beaver Biosciences Inc. (Suzhou, China). Taq PCR Master Mix and ChamQ Universal SYBR qPCR Master Mix were purchased from Vazyme Biotechnology Co., Ltd. (Nanjing, China). The 96-well polystyrene microtiter plate (MaxiSorp) was bought from Thermo Fisher Scientific (Shanghai, China). ASFV-positive freeze-dried serum, porcine negative serum national reference was purchased from the National Center of Veterinary Culture collection (CVCC) (Beijing, China).
4
FITC-Protein Binding Protocol
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All reagents were of analytical grade, unless mentioned otherwise. Amino acids were purchased from Kangda Amino Acid Works (Shanghai, China). Fluorescein isothiocyanate (FITC), polyvinylpyrrolidone and all proteins were obtained from Sigma-Aldrich (St. Louis, USA). 5 × TBE buffer (0.225 M Tris/boric acid, 0.05 M EDTA) for DNA separation was purchased from Sangon Biotech (Shanghai, China). SYBR Green I used as a double-stranded DNA binding fluorescent dye was purchased from Xiamen Zeesan Biotech (Xiamen, China). FastDigest Mva I for DNA digestion was purchased from Thermo Fisher Scientific (Waltham, USA). A 200 bp DNA fragment as an internal standard in DNA separation was obtained from Beijing Biomed (Beijing, China), and other reagents for DNA separation were purchased from Sinopharm Chemical Reagent (Shanghai, China). All of aqueous solutions were prepared within deionized water, then filtered through 0.22 μm Millipore filters (Darmstadt, Germany), and stored at 4 °C.
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