Kit for examining Liddle's related gene mutation
A kit and gene technology, applied in the field of molecular biology, can solve the problems of low sensitivity and specificity, strict detection time requirements, unbearable cost, etc., to avoid uncertain primers, improve detection accuracy and stability, increase Effects on Clinical Detection Rate and Accuracy
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[0047] 1. Probe design:
[0048] Synthesize streptomycin-labeled probes according to the table. The probe solution uses the SureSelect buffer system of agilent company.
[0049] 2. Genome extraction:
[0050] Qiagen FlexiGene DNA Kit (Code No: 51204) was used to extract the genomes of 96 samples to be tested, OD 260 / 280 When the value reaches 1.8-2.0, take 1-3g each as the starting template.
[0051] 3. Sample preparation before sequencing
[0052] 1. Target gene fragmentation:
[0053] Take the quantified 270 copies of genomic DNA and dilute to 20-35ng / L. Take 130L and fragment them separately with a sonicator.
[0054] 2. AMpure XP fragment selection
[0055] 1. Take a 96-well plate, add 80-100L magnetic beads, then add sample DNA, pipette repeatedly to mix;
[0056] 2. Place the mixture at 26°C for 5 minutes;
[0057] 3. Put the 96-well plate on the magnetic plate for 3 minutes;
[0058] 4. Carefully pipette all the supernatant into a new well, be careful not to t...
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