A kind of nucleic acid encapsulation reagent and its application
A nucleic acid and reagent technology, which is applied in the field of nucleic acid encapsulation reagents, can solve the problems of time-consuming and labor-intensive, low success rate, etc., and achieve good dissolution effect, good adhesion effect, and good nucleic acid embedding effect
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Embodiment 1
[0060] Embodiment 1, the preparation of packaging reagent
[0061] Encapsulation reagent: composed of 5 parts by mass of agarose, 3 parts by mass of saponin and 4 parts by mass of sodium hydroxypropyl cellulose.
Embodiment 2
[0062] Embodiment 2, preparation for detecting the reaction device of Zika virus
[0063] 1. Mix the encapsulation reagent prepared in Example 1, primer ZIKV-F, primer ZIKV-R, probe ZIKV-P and sterile water to obtain embedding solution. In the embedding solution, the concentration of the encapsulation reagent was 0.1 g / 100 ml, the concentration of the primer ZIKV-F was 2 μM, the concentration of the primer ZIKV-R was 2 μM, and the concentration of the probe ZIKV-P was 1 μM.
[0064] Primer ZIKV-F: 5'-TGCATWGGAGTCAGCAA-3';
[0065] Primer ZIKV-R: 5'-AATTCTAATACGACTCACTATAGGGAGATAGGATCTTACTCVGCCA-3';
[0066] Probe ZIKV-P: 5'-FAM-CGGACGTCAGGTGGGACYTGGGTTGATGTTGTCTTGGGCCTG-BHQ2-3'.
[0067] The primer ZIKV-F is shown as sequence 1 in the sequence listing. The primer ZIKV-R is shown as sequence 2 in the sequence listing. The nucleotide sequence of the probe ZIKV-P is shown as sequence 3 in the sequence listing, the 5' end has a fluorescent group FAM, and the 3' end has a quenc...
Embodiment 3
[0070] Embodiment 3, the preparation of the kit for detecting Zika virus
[0071] 1. Preparation of kits
[0072] The kit consists of the following components (each component is individually packaged):
[0073] (1) The reaction device for detecting Zika virus prepared in Example 2.
[0074] (2) Amplification reaction solution BQ: the solvent is Tris-HCl buffer solution with pH 8.0 and 100 mM; the solute and its concentration are as follows: 2 mM dNTP (that is, the concentration of dATP, dTTP, dCTP and dGTP is 2 mM), 4 mM rNTP ( That is, the concentrations of rATP, rTTP, rCTP and rGTP are all 4mM), 50mM DTT, 60mM MgCl 2 , 300mM KCl, 15% (volume percentage content) DMSO, 1M sorbitol.
[0075] (3) Enzyme mixture EQ: the solvent is water; the solute and its concentration are as follows, AMV reverse transcriptase 0.8U / μl, T7 RNA polymerase 3.5U / μl, ribonuclease H 0.3U / μl, pyrophosphatase 0.5U / μl μl, RNase inhibitor 5U / μl, BSA 0.5μg / μl.
[0076] (4) Positive quality control pro...
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