Method and kit for transforming and purifying DNA in DNA methylation detection process
A detection process and methylation technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as DNA fragmentation, false positive DNA methylation detection results, and difficulty in mechanization, and improve detection efficiency. , The effect of saving detection energy consumption and saving detection cost
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Embodiment 1
[0026] In order to achieve the above object, the method steps of DNA conversion and purification in the DNA methylation detection process ( figure 1 )for:
[0027] 1) In the supersaturated sodium bisulfite / sodium hydroxide solution, add the DNA sample without adding any DNA protecting agent;
[0028] 2) Carry out DNA transformation at 70°C, and the transformation time is 20 minutes;
[0029] 3) After step 2) is completed, add an appropriate amount of 2mol / L citric acid + 3mol / L sodium citrate + 6mol / L guanidine hydrochloride, and use magnetic beads to absorb the converted DNA;
[0030] 4) For the DNA adsorbed by the magnetic beads in the step 3), the purified DNA is obtained by washing with 8mol / L sodium perchlorate+dehydrated ethanol, isopropanol and tris(hydroxymethyl)aminomethane-hydrochloric acid solution respectively;
[0031] 5) The purity and concentration of the purified DNA were tested, which met the requirements for further DNA methylation detection.
Embodiment 2
[0033] In order to achieve the above object, the method steps of DNA conversion and purification in the DNA methylation detection process ( figure 1 )for:
[0034] 1) In supersaturated sodium sulfite / ammonium sulfite monohydrate / sodium bisulfite solution, add DNA sample without adding any DNA protectant;
[0035] 2) Carry out DNA transformation at 70°C, and the transformation time is 20 minutes;
[0036] 3) After step 2) is completed, add an appropriate amount of absolute ethanol + 3mol / L sodium chloride + 1mol / L tris(hydroxymethyl)aminomethane-hydrochloric acid solution, and use magnetic beads to absorb the transformed DNA;
[0037] 4) For the DNA adsorbed by the magnetic beads in step 3), wash with 1 mol / L sodium chloride + 0.5% sodium lauryl sulfate + isopropanol, absolute ethanol and nucleic acid-free water respectively to obtain Purified DNA;
[0038] 5) The purity and concentration of the purified DNA were tested, which met the requirements for further DNA methylation...
Embodiment 3
[0040] In order to achieve the above object, the method steps of DNA conversion and purification in the DNA methylation detection process ( figure 1 )for:
[0041] 1) In supersaturated sodium bisulfite / ammonium bisulfite monohydrate / ammonium bisulfite solution, add DNA sample without adding any DNA protectant;
[0042] 2) Carry out DNA transformation at 70°C, and the transformation time is 20 minutes;
[0043] 3) After step 2) is completed, add an appropriate amount of 2mol / L citric acid + 4mol / L guanidine hydrochloride solution, and use magnetic beads to absorb the transformed DNA;
[0044] 4) For the DNA adsorbed by magnetic beads in step 3), pass through 1mol / L sodium chloride+sodium lauryl sulfate+absolute ethanol with a mass percentage of 0.5%, isopropanol and tris(hydroxymethyl)aminomethane respectively - washing with hydrochloric acid solution to obtain purified DNA;
[0045] 5) The purity and concentration of the purified DNA were tested, which met the requirements ...
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