Method for detecting neomycin B content in milk and application thereof
A technology of neomycin B and neomycin, applied in biochemical equipment and methods, biological testing, material inspection products, etc., can solve the problems of slow analysis speed, high detection cost, expensive detection instruments, etc., and achieve easy operation , highly specific effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0040] Embodiment 1, the preparation of the nucleic acid aptamer that detects neomycin B (NeomycinB) in milk
[0041] 1. Preparation of nucleic acid aptamers
[0042] The RNA aptamer (rGrGrArCrUrGrGrGrGrCrGrArGrArArGrUrUrUrArGrGrUrCrC, wherein rG represents the G on the RNA) in the sequence capable of binding to neomycin B is truncated into two parts in its segment rA and rG, and one part is in its sequence The 3' end is modified with 15 thymine (T) spacers and 12 adenine (A) tails, named NEO1, and its nucleotide sequence is as follows: 5'-rGrArArGrUrUrUrArGrUrCrC-(T) 15 -(A) 12 -3' (sequence 1); another part modifies chemical fluorescent group (FAM) at its sequence 5' end, named after (NEO2), and its nucleotide sequence is 5'-FAM-rGrGrArCrUrGrGrGrGrCrGrA-3' (sequence 2 ).
[0043] Artificially synthesized NEO1 and NEO2 are the nucleic acid aptamers of neomycin B.
[0044] 2. Preparation of a kit for detecting neomycin B (NeomycinB) in milk
[0045] Preparation of nano-go...
Embodiment 2
[0053] Embodiment 2, detection of neomycin B (NeomycinB) in milk
[0054] 1. Sensitivity detection
[0055] The milk to be tested contains 250mMNaCl and 5mMMgCl 2 The 50mM Tris-HCl buffer solution was diluted 1:5 to obtain the test sample; stored at 4°C.
[0056] Take 284 μL of 10 nM nano-gold colloid solution linked to NEO1, 6 μL of 100 μM NEO2 solution, and 10 μL of detection sample prepared in Example 1 above, mix them and let stand at room temperature for 10 minutes to obtain a reaction solution.
[0057] Detect the fluorescence value of the reaction solution with a fluorescence detector; calculate the neomycin B content by the fluorescence value of the reaction solution;
[0058] The calculation formula is as follows: fluorescence value=-1.331×concentration+56.760.
[0059] The result is as figure 2 As shown, it can be seen that the concentration of neomycin B was detected linearly within 0.1 μM to 10 μM, and the lowest detection limit was 0.001 μM.
[0060] 2. Spec...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com