CARD9 gene mutant and application thereof
A mutant, c.191instgct technology, used in applications, genetic engineering, plant genetic improvement, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0087] Example 1 Exon sequencing to determine the mutation site
[0088] Select three cases (D6-1, D6-3, D6-4 (wherein, D6-1 and D6-3 are patients from Peking University First Hospital; D6-4 is a patient from Shandong Qilu Hospital)) confirmed Perform exome sequencing and data analysis for sporadic patients with phaeohyphomycosis. The specific procedures are as follows:
[0089] First, the inventors sequenced the exome sequences of the three patients using NimbleGen 2.1M Human Exome Array combined with Solexa high-throughput sequencing technology, as follows:
[0090] 1) The genomic DNA of the patient sample was randomly broken into fragments of about 200-300 bp, and then according to the operating instructions provided by the manufacturer, adapters were connected to both ends of the fragments to prepare a hybrid library. After purification, the library undergoes linear amplification of Linker Mediated PCR (LM-PCR) and hybridization enrichment with Biotinylated DNA Library, a...
Embodiment 2
[0095] Example 2 Sanger method sequencing verification
[0096] 3 patients (D6-1, D6-3, D6-4), 6 normal people in the family (that is, the parents of the 3 patients, none of them had the disease), and a sporadic patient (from Wuhan No. Hospital) gene detection, design primers for all exon sequences of CARD9 gene, and then obtain CARD9-related sequences by PCR amplification, product purification and sequencing, and verify whether CARD9 is mutant or wild-type according to the sequence determination results Association with phaeohyphomycosis disease. The specific method steps are as follows:
[0097] 2.1 DNA extraction
[0098] The peripheral blood of the above-mentioned 4 patients with phaeohyphomycosis and 6 normal people in the family were collected respectively, and then the genomic DNA in the peripheral blood leukocytes was extracted by the conventional phenol-chloroform method, and the concentration of the extracted DNA was measured by a spectrophotometer and purity, the...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 