The total RNA of the cryopreservation samples from the refrigerator at −80 °C was extracted using an EASYspin Plus Complex Plant RNA kit (Aidlab, China), referring to the manufacturer’s instructions, and DNA was eliminated with DNase I (Aidlab, China). The purity and concentration of RNA samples were detected using 1% agarose gel electrophoresis and a Synergy H1 multifunction microplate detector (BioTek, American), respectively. Additionally, 0.2 μg of total RNA with a 260/280 ratio between 1.9 and 2.1 was used to synthesize first-strand cDNA using HiScript® II Q RT SuperMix for qPCR (Vazyme, Nanjing, China) in accordance with the manufacture’s manual, and then the sample was diluted five times for qPCR analyses.
Dnase 1
Manufactured by Aidlab
Sourced in China
DNase I is a digestive enzyme that catalyzes the hydrolytic cleavage of DNA. It is a versatile tool commonly used in molecular biology and genetic research applications.
Automatically generated - may contain errors
Lab products found in correlation
Easyspin plus complex plant rna kit, by Aidlab (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Hiscript 2 q rt supermix for qpcr, by Vazyme (1 mentions)
Fastquant rt kit, by Tiangen Biotech (1 mentions)
Plant total rna extraction kit, by Aidlab (1 mentions)
Nanodrop 2000c spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Goldenstar rt6 cdna synthesis kit, by Tsingke (1 mentions)
3 protocols using dnase 1
1
Isolation and Quantification of Plant RNA
2
RNA Extraction and qRT-PCR Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total RNA was isolated from the collected materials with a plant total RNA extraction kit (Aidlab, Beijing, China) according to the manufacturer's protocol and treated with DNase I (Aidlab, Beijing, China). A NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, West Palm Beach, FL) was used to measure RNA quality and quantity. Approximately 2 μg total RNA was used for reverse transcription with a Tiangen Fast Quant RT Kit (Tiangen Biotech Co. Ltd., Beijing, China) according to the manufacturer's instructions. The qRT‐PCR was conducted as previously described (Bustin et al., 2009 ; He et al., 2018 ). Primer Premier v.6 (Sigma‐Aldrich Corp., St. Louis, MO) was used to develop the primers listed in Table S2 . At least 20 replicates (five biological replicates × four technical replicates) were performed per experiment.
3
RNA Extraction and cDNA Synthesis
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Total RNA was isolated from collected samples using the EASYspin Plus Complex Plant RNA kit (catalog number: RN53, Aidlab Biotechnologies Co., Ltd, Beijing, China) according to the manufacturer's instructions and treated with DNase I (catalog number: RN34, Aidlab, Beijing, China) to eliminate DNA contamination. The RNA concentration and purity were determined with a NanoDrop 2000c Spectrophotometer (Thermo Scientific, US), and RNA integrity was checked on 1% agarose gel electrophoresis. The extracted RNA had a 260/280 ratio between 1.9 and 2.1, which was considered to meet the requirements of following experiments. 0.6 μg of total RNA was prepared for synthesizing first-strand of cDNA by the Goldenstar™ RT6 cDNA Synthesis kit (Tsingke, Nanjing, China) according to the manufacture's instructions.
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