A kit for high-throughput detection of natural water samples of Prorocentrum donghaiense
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, 1.4M NaCl, 20mM EDTA, 100mMTris-Cl, 10URNase Inhibitor;
[0029] (2) Reagent B is 5×NASBABuffer, its composition is 0.2MTris-HCl, 60mMMgCl 2 ,0.35MKCl 2 , 25mM DTT, 20mMdNTP;
[0030] (3) Reagent C is NTP (10mMeach);
[0031] (4) Reagent D is 5×Primermix, its composition is: 75%DMSO, 1μM NASBA-P1, 1μM NASBA-P2, 25mMDTT, 20mMdNTP;
[0032] (5) Reagent E is 4×Enzymemix, its composition is: 1.5M sorbitol, 0.42mg / mlBSA, 16U / mlRNaseH, 6400U / mlT7RNAPolymerase, 1280U / mlRT-M-MLV, 4000U / mlRNA inhibitor;
[0033] (6) Reagent F is RNA denaturant, its composition is: 1.5M NaCl, 0.15M trisodium citrate dihydrate, pH7.0, 0.15M formaldehyde;
[0034] (7) Reagent G is hybridization solution, its compos...
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