All oligonucleotide sequences were synthesized and purchased from Sangon Inc. (Shanghai, China). Table S1 in ESI† shows the sequences of the oligonucleotides used in the study. All oligonucleotides were HPLC-purified and dissolved in tris-ethylenediaminetetraacetic acid (TE) buffer (pH 8.0, 10 mM Tris–HCl, 1 mM ethylene diamine tetraacetic acid) and stored at −20 °C, which were diluted in appropriate buffer prior to use. DL20 DNA marker was purchased from Takara (Dalian, China). Gold view (GV) was purchased from SBS Genetech (Beijing, China). Klenow fragment and Nb.BbvCI nicking enzyme were purchased from New England Biolabs Inc. (Beverly, MA, USA). Deoxynucleotide triphosphates (dNTPs) were obtained from Sangon Inc. (Shanghai, China). All other chemicals not mentioned here were of analytical reagent grade. Millipore-Q water (≥18 MΩ) was used in all experiments. Human serum samples were obtained from the First Affiliated Hospital of Chongqing Medical University.
Nb bbvci nicking enzyme
Manufactured by New England Biolabs
Sourced in United States
Nb.BbvCI is a nicking enzyme that introduces a single-strand nick in DNA. Its core function is to generate a site-specific nick in double-stranded DNA.
Automatically generated - may contain errors
2 protocols using nb bbvci nicking enzyme
1
Oligonucleotide-based Assay Development
2
Fluorescent DNA Methylation Assay
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Dam and M.SssI MTase, Dpn I, Nb.BbvCI nicking enzyme, Klenow Fragment and S-adenosylmethionine (SAM) were gotten from New England Biolabs (Ipswich, MA, USA). Gentamycin, benzyl penicillin and all oligonucleotides (Table 1) used in this study were purchased from Sangon Biotechnology Co., Ltd. (Shanghai, China). All other reagents were of analytical reagent grade (AR), and without purification for usage. Ultrapure water with 18.2 MΩ•cm resistivity was used to prepare the solutions.
A Hitachi F-7000 fluorescence spectrometer (Hitachi Ltd., Japan) was selected for all fluorescence measurements of sample solutions. The emission spectra scope ranged from 500 to 600 nm, while the wavelength of excitation light was fixed at 480 nm under room temperature. At 520 nm, the fluorescence was measured as the emission intensity. The voltage of the photomultiplier tube was set at 700 V and the emission slit was 5 nm, the same as the excitation slit.
A Hitachi F-7000 fluorescence spectrometer (Hitachi Ltd., Japan) was selected for all fluorescence measurements of sample solutions. The emission spectra scope ranged from 500 to 600 nm, while the wavelength of excitation light was fixed at 480 nm under room temperature. At 520 nm, the fluorescence was measured as the emission intensity. The voltage of the photomultiplier tube was set at 700 V and the emission slit was 5 nm, the same as the excitation slit.
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