Nucleotide sequence and method for identifying goat milk components and mutton bone meal components in feed
A technology of mutton and bone meal, applied in the direction of DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of inability to identify and distinguish the source of component tissues, low specificity, narrow application range, etc., and achieve easy promotion, high sensitivity, Good species-specific effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0054] Example 1 Selection of target sequence of sheep tissue components and design and synthesis of primers
[0055] In order to identify goat milk-derived ingredients and prohibited goat-derived ingredients, it is necessary to find a tissue-specific marker as a detection marker, which should have the following characteristics:
[0056] (1) The marker can be detected in bone, muscle, blood, and internal organs;
[0057] (2) The marker cannot be detected in milk;
[0058] (3) The marker has a certain stability, that is, it can still be detected after the bone, muscle and viscera are processed into meat and bone meal;
[0059] (4) It is conserved within a species and has specificity among species.
[0060] According to the mRNA fragments of sheep osteonectin recorded in Genbank, three pairs of primers (YG51, YG52, YG53) were designed for the fragments of sheep osteonectin mRNA, which were compared and analyzed with DNAMAN software. In order to avoid the interference of genomi...
Embodiment 2
[0065] Example 2 Extraction of sample total RNA
[0066] Use the RNA extraction kit from TIANGEN company to extract total RNA, the specific operation is as follows:
[0067] (1) Take 25 mg of sample, add 1 ml of lysate RZ, shake and mix well.
[0068] (2) Leave the sample at room temperature for 5 minutes. Centrifuge at 12,000 rpm at 4°C for 5 minutes, take the supernatant, and transfer it to a new RNase-free centrifuge tube.
[0069] (3) Add 200 μl of chloroform, cover the tube cap, shake vigorously for 15 seconds, and place at room temperature for 3 minutes.
[0070] (4) Centrifuge at 12000rpm at 4°C for 10 minutes. The sample is divided into three layers: a yellow organic phase, an intermediate layer and a colorless aqueous phase. RNA is mainly in the aqueous phase, and the volume of the aqueous phase is about 50% of the lysate RZ reagent used. . Transfer the aqueous phase to a new tube.
[0071] (5) Slowly add 1.5 times the volume of absolute ethanol and mix well. Th...
Embodiment 3
[0078] Example 3 Establishment of real-time fluorescent RT-PCR detection method
[0079] Choose TaKaRa's The RT-PCR kit is used for RT-PCR amplification. Through repeated trials, the reaction system and reaction conditions are optimized to achieve the best experimental results.
[0080] The final determination of the reaction system is as follows:
[0081]
[0082] SYBR Real Time RT-PCR reaction conditions are set as follows:
[0083]
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