Kit capable of detecting natural water sample of prorocentrum donghaiense in high throughput manner
A technology of Prorocentrum and kit, applied in the field of kits capable of high-throughput detection of natural water samples of Prorocentrum in the East China Sea
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[0026] The present invention will be described in detail below in conjunction with specific embodiments.
[0027] 1. Kit composition
[0028] (1) Reagent A is a crude nucleic acid extraction reagent for natural samples, and its composition is: 2% (m / v) CTAB, 0.5% (v / v) β-mercaptoethanol, 1.4 M NaCl, 20 mM EDTA, 100 mM Tris-Cl , 10 U RNase Inhibitor;
[0029] (2) Reagent B is 5×NASBA Buffer, its composition is 0.2 M Tris-HCl, 60 mM MgCl 2 , 0.35 M KCl 2 , 25 mM DTT, 20 mM dNTP;
[0030] (3) Reagent C is NTP (10 mM each);
[0031] (4) Reagent D is 5×Primer mix, its composition is: 75% DMSO, 1 μM NASBA-P1, 1 μM NASBA-P2, 25 mM DTT, 20 mM dNTP;
[0032] (5) Reagent E is 4×Enzyme mix, its composition is: 1.5 M sorbitol, 0.42 mg / ml BSA, 16 U / ml RNase H, 6400 U / ml T7 RNA Polymerase, 1280 U / ml RT-M-MLV , 4000 U / ml RNA inhibitor;
[0033] (6) Reagent F is RNA denaturant, its composition is: 1.5 M NaCl, 0.15 M trisodium citrate dihydrate, pH 7.0, 0.15 M formaldehyde;
[0034] (...
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