Method and kit for non-invasive measurement on fetus deaf pathogenic gene mutation
A disease-causing gene and kit technology, applied in the detection of fetal deafness-causing gene mutations, non-invasive detection of fetal deafness-causing gene mutations, can solve the problem that no one can achieve non-invasive fetal deafness-causing gene detection
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[0133] Experimental steps:
[0134] 1. Plasma DNA Extraction
[0135] Take 1-2mL of plasma, and use the QIAampCirculatingNucleicAcidKit Cat. No. 55114 to extract plasma DNA. Finally, DNA was eluted with 45 μl of elution buffer, and 2 μl of Qubit was used to measure the concentration.
[0136] 2. Plasma DNA filling plus A
[0137] Prepare the reaction mixture in Table 1:
[0138] Table 1
[0139] T4 DNA Polymerase Buffer (10X)
5μl
40.5μl
Taq
0.5μl
2.0μl
10mM dNTPs
2.0μl
total capacity
50μl
[0140] Reaction on the PCR machine:
[0141] 37°C: 20min
[0142] 72°C: 20min
[0143] 4°C: keep
[0144] The product added with A was purified by column, dissolved in 25 μl BufferEB, and eluted twice.
[0145] 3. Joint connection
[0146] Prepare the reaction mixture in Table 2:
[0147] Table 2
[0148] dna
22μl
2x Quick Ligase Buffer
25μl
...
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