Method of authenticating chicken contour feathers and silken feathers by applying fluorescence probe PCR (polymerase chain reaction)
A fluorescent probe and identification method technology, which is applied in the field of chicken molecular marker preparation, can solve the problems of high production cost and long breeding time, and achieve the effects of short detection time, strong specificity and high detection sensitivity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0037] Example 1 : Design and synthesis of primers and Taqman probes
[0038] A pair of specific primers and two Taqman probes (Table 1) were designed by using the PrimerExpress 5.0 software according to the DNA sequence of chicken chromosome 3 in Ganbank and chicken feather traits.
[0039] Table 1 Primer and Taqman probe sequence (5'-3')
[0040]
Embodiment 2
[0041] Example 2: Fluorescent probe PCR detection
[0042] 2.1 Extraction of nucleic acid
[0043] The DNA of GG genotype, CC genotype and GC genotype was extracted according to the instructions of the blood genome column extraction kit.
[0044] 2.2 Preparation of standard genotypes
[0045] PCR amplification was performed with self-designed primers, the upstream primer was 5'-GACTCTCAACGCGGGAAC-3', and the downstream primer was 5'-CTGGGGGCAGCCATCTT-3'.
[0046] A 25 μL reaction system was used: 2×EsTaqMasterMix 15 μL, forward and reverse primers 0.5 μL each, DNA template 2 μL, and water to 25 μL.
[0047] The PCR conditions are: first step: 95°C for 5min, second step: 95°C for 30s, third step: 62°C for 30s, fourth step: 72°C for 30s, fifth step: 72°C for 5min; Four-step cycle 30 times.
[0048] The PCR product was detected by 1% agarose gel electrophoresis, and the target gene was recovered and ligated with pGM-Tvector to obtain a recombinant plasmid.
[0049] The lig...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 