Probe library and kit for detecting lung cancer related 41 genes and application thereof
A kit and technology for lung cancer, applied in the field of biomedicine, can solve the problems of not being able to cover the spread of genes, and achieve the effects of good sequencing depth uniformity, maximum use, and reduced clinical testing costs
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Embodiment 1
[0201] 1. Build a sample library
[0202] 1. Extraction of DNA
[0203] Sample DNA was extracted using the HiPure FFPE DNA Kit (Magen) kit.
[0204] 2. DNA Fragmentation
[0205] According to the instruction manual of the DNA breaker (Bandelin), the DNA sample was fragmented so that the fragment length was 200-500 bases.
[0206] DNA was purified using the Beckman Coulter Ampure Beads kit (Cat. No. A63881).
[0207] 3. DNA sample bank quality inspection
[0208] Use Bio-Fragment Analyzer to conduct qualitative and quantitative analysis of DNA to confirm that the peak length of DNA fragments is reasonable.
[0209] 4. DNA end repair
[0210] Using T4 polymerase and Klenow E. coli polymerase fragments, blunt the 5' protruding sticky ends and 3' protruding sticky ends of DNA to generate blunt ends. At the same time, T4 polynucleotide kinase is used to phosphorylate the 5' end of DNA for subsequent blunt end ligation.
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[0213] The reaction was carrie...
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