US 12,442,049 B2
Method for rapidly detecting the total nitrogen content of the soil in an urban green land by using archaea molecular marker OTU300
Jigang Han, Shanghai (CN); Lang Zhang, Shanghai (CN); Weiwei Zhang, Shanghai (CN); and Gangyong Zhao, Shanghai (CN)
Assigned to SHANGHAI ACADEMY OF LANDSCOPE ARCHITECTURE SCIENCE AND PLANNING, Shanghai (CN)
Appl. No. 17/595,214
Filed by Shanghai Academy Of Landscape Architecture Science And Planning, Shanghai (CN)
PCT Filed May 14, 2020, PCT No. PCT/CN2020/090172
§ 371(c)(1), (2) Date Nov. 11, 2021,
PCT Pub. No. WO2021/142989, PCT Pub. Date Jul. 22, 2021.
Claims priority of application No. 202010062417.6 (CN), filed on Jan. 19, 2020.
Prior Publication US 2022/0220538 A1, Jul. 14, 2022
Int. Cl. C12Q 1/6851 (2018.01); C12Q 1/689 (2018.01); G01N 33/24 (2006.01)
CPC C12Q 1/689 (2013.01) [C12Q 1/6851 (2013.01); G01N 33/24 (2013.01)] 3 Claims
OG exemplary drawing
 
1. A method for detecting a total nitrogen content in soil comprising the following steps:
providing a primer comprising the sequence of SEQ ID NO. 4 and a primer comprising the sequence of SEQ ID NO. 5 as an amplification primer pair;
providing a nucleotide sequence of a probe shown as SEQ ID NO. 1 of the sequence listing;
using the isolated total DNA of a soil sample as a template to perform real-time fluorescent quantitative PCR on the soil sample on a fluorescent quantitative PCR instrument; and
obtaining a cycle threshold (Ct) value by real-time fluorescent quantitative PCR and calculating a copy number according to the Ct value, and calculating the total nitrogen content by the copy number content in the soil sample;
wherein the probe is a real-time fluorescent quantitative PCR probe with a fluorescent group 5(6)-Carboxyfluorescein at its 5′ end and a fluorescence quenching group 5(6)-Carboxytetramethylrhodamine at its 3′ end.