Lysis solution for extracting nucleic acid through magnetic bead method
A technology for extracting nucleic acid and lysate, which is applied in the field of molecular biology, can solve the problems of low efficiency of lysate, and achieve the effect of authentic and reliable experimental results, good dispersion, and effective cell lysis
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Embodiment 1
[0063] Embodiment 1: The real-time fluorescent quantitative PCR detection of HBVDNA
[0064] The reagents prepared according to the above-mentioned lysate scheme 1) and washing solution scheme 1) mixed with magnetic beads were used respectively for the clinical diagnosis and the tested HBVDNA quantitative value of 2×10 1 IU / ml hepatitis B patient's serum carries out the operation of following steps:
[0065] (1) Dispensing of PCR lysate: Dispense the prepared lysate mixed with magnetic beads into dedicated PCR tubes at a rate of 100 μl per tube.
[0066] (2) Adding samples: Take 100 μl of serum and add it to the above-mentioned PCR tube containing the lysate mixed with magnetic beads, gently blow and mix 5 times with a tip, and let stand at room temperature for 10 minutes.
[0067] (3) Aspirate and discard the liquid: place the above-mentioned PCR tubes on the eight-row magnetic stand, let it stand for 2 minutes, and use a pipette to absorb the liquid on the opposite side of ...
Embodiment 2
[0075] Embodiment 2: The real-time fluorescent quantitative PCR detection of HBVDNA
[0076] The reagents prepared according to the above-mentioned lysate scheme 2) and washing solution scheme 2) mixed with magnetic beads were used respectively for the clinical diagnosis and the tested HBVDNA quantitative value of 2×10 1 IU / ml hepatitis B patient's serum carries out the operation of following steps:
[0077] (1) Dispensing of PCR lysate: Dispense the prepared lysate mixed with magnetic beads into dedicated PCR tubes at a rate of 100 μl per tube.
[0078] (2) Adding samples: Take 100 μl of serum and add it to the above-mentioned PCR tube containing the lysate mixed with magnetic beads, gently blow and mix 5 times with a pipette tip, and let stand at room temperature for 5 minutes.
[0079] (3) Aspirate and discard the liquid: place the above-mentioned PCR tubes on the eight-row magnetic stand, let it stand for 2 minutes, and use a pipette to absorb the liquid on the opposite s...
Embodiment 3
[0087] Embodiment 3: The real-time fluorescent quantitative PCR detection of HBVDNA
[0088] Prepare the lysate and wash solution mixed with magnetic beads according to the following component ratios:
[0089] 0.3N Sodium Hydroxide, 0.45M Potassium Chloride, 0.03% Sodium N-Lauroyl Sarcosinate, 5mM EDTA, 0.45M Tris-HCL, 1% Triton X-100, 3mMDTT and 0.001wt% Bromophenol Blue.
[0090] Add the magnetic beads that have naturally precipitated for 24 hours according to the volume ratio of magnetic beads and lysate of 1:100, and mix well.
[0091] Washing liquid: 0.3M potassium chloride, 0.001wt% stone green, adjust the pH value to 4 with acetic acid.
[0092] Use the above lysate and wash solution to confirm the clinical diagnosis and the tested HBVDNA quantitative value is 2×10 1 IU / ml hepatitis B patient's serum carries out the operation of following steps:
[0093] (1) Dispensing of PCR lysate: Dispense the prepared lysate mixed with magnetic beads into dedicated PCR tubes at a...
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