Specific primers and typing method of class I mhc gene for the detection of antiviral potential of crested ibis
A genotyping method and technology of a genotyping method, applied in the field of genotyping, can solve the problems of low reproductive ability, serious inbreeding, high death and disability rate of young birds, etc.
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example 1
[0044] 1. Sample preparation: Crested ibis blood and tissue samples came from artificial populations such as Deqing, Sado, Beijing, Henan, Yangxian and Louguantai.
[0045] 2. DNA extraction: phenol-chloroform method to extract blood and tissue DNA.
[0046] 3. Primer synthesis: According to the obtained crested ibis MHC gene sequence, design universal primers to amplify exon 2 for the typing of UAA loci, and BGI synthesizes primers. The primer sequences are as follows:
[0047]
[0048] 4. Primer amplification: the above-mentioned amplification primers are respectively applied to PCR to amplify the extracted sample DNA. Set the PCR thermal cycle annealing temperature range to 61°C.
[0049] 5. Amplification result detection: take 2 ul of the above-mentioned PCR amplification products and carry out agarose gel electrophoresis with a gel concentration of 1.5%. The result is as figure 1 As shown, this primer can effectively amplify the target band, and the fragment size o...
example 2
[0063] 1. Sample preparation: Crested ibis blood and tissue samples came from artificial populations such as Deqing, Sado, Beijing, Henan, Yangxian and Louguantai.
[0064] 2. DNA extraction: phenol chloroform method to extract blood and tissue DNA.
[0065] 3. Primer synthesis: Design specific primers for exon 3 of the UBA site based on the obtained Crested Ibis MHC gene sequence, and BGI synthesized the primers. The primer sequences are as follows:
[0066]
[0067] 4. Primer amplification: apply the above amplification primers to PCR respectively to amplify the extracted sample DNA. Set the PCR thermal cycle annealing temperature range to 61°C.
[0068] 5. Amplification result detection: take 2 ul of the above-mentioned PCR amplification products and carry out agarose gel electrophoresis, the gel concentration is 1.5%. The result is as figure 2 As shown, this primer can effectively amplify the target band, and the fragment size of the target band is 150 bp. The PCR ...
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