Magnetic bead mixture, nucleic acid preservation method, nucleic acid extraction reagent and kit
A mixed liquid and magnetic bead technology, applied in the field of biochemistry, can solve the problems of inability to store at room temperature or refrigeration, inability to judge DNA fragmentation, complicated preparation, etc., to reduce packaging costs and transportation costs, save sample addition steps, Simple to use effects
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Embodiment 1
[0055] 1. The nucleic acid extraction steps are as follows:
[0056] (1) Add 450 μL of cell lysate, 40 μL of magnetic bead mixture, and 250 μL of sample into a 1.5 mL centrifuge tube, mix well for 2 minutes, and then let stand at room temperature for 10 minutes. Perform a brief centrifugation to absorb the magnetic beads and discard all the solution.
[0057] (2) Add 500 μL of washing solution, mix for 2 minutes, and then let stand at room temperature for 2 minutes. Perform a brief centrifugation to absorb the magnetic beads and discard all the solution.
[0058] (3) Add 500 μL of washing solution, mix for 2 minutes, and then let stand at room temperature for 2 minutes. Perform a brief centrifugation to absorb the magnetic beads and discard all the solution.
[0059] (4) Dry the magnetic beads at room temperature for 10 minutes, add 80 μL of eluent, mix for 2 minutes, and then let stand at room temperature for 5 minutes. Perform brief centrifugation to absorb the magnetic ...
Embodiment 2
[0083] The nucleic acid extraction step and fluorescent quantitative PCR detection step of Example 1 were used to analyze the recovery rate of the magnetic bead mixture.
[0084] The magnetic bead mixed liquid formula of the present embodiment is as described in table 6:
[0085] Table 6
[0086] component name Dosage range per mL magnetic beads 100mg Proteinase K 50mg urea 10mM Tris-Hcl 100mM CaCl 2
10mM EDTA-Na 2
10mM glycerin 50% Carrier RNA 25μg DNase and RNase free purified water Replenish to 1mL
[0087] Detection of the recovery rate of the magnetic bead mixture:
[0088] a) Use negative serum to dilute the recombinant plasmid to 5.6×10 4 copy / μL, the magnetic bead mixture was used for extraction, and the PCR reaction reagent was used for detection.
[0089] b) Dilute the recombinant plasmid to 1.75×10 with DNase / RNase-free purified water 5 copy / μL, detected by PCR reaction reagent...
Embodiment 3
[0099] The nucleic acid extraction step and fluorescent quantitative PCR detection step of Example 1 were used to analyze the recovery rate of the magnetic bead mixture.
[0100]The magnetic bead mixed liquid formula of the present embodiment is as described in table 8:
[0101] Table 8
[0102]
[0103]
[0104] Detection of the recovery rate of the magnetic bead mixture:
[0105] a) Dilute the recombinant plasmid to 1×10 with negative serum 5 copy / μL, 1×10 4 copy / μL, 1×10 3 copy / μL, the magnetic bead mixture is used for extraction, and the reaction reagent is used for detection.
[0106] b) Use DNase / RNase-free purified water to dilute the recombinant plasmid to 3.125×10 5 copy / μL, 3.125×10 4 copy / μL, 3.125×10 3 copy / μL, directly detected by the reaction reagent.
[0107] Table 9
[0108]
[0109]
[0110] c) The results are shown in Table 9 and image 3 As shown, after the mixed recombinant plasmid serum was extracted, 1×10 5 copy / μL (equivalent to 3....
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