Rice tissue samples (50–100 mg) were ground using a TisseLyser II (Quiane Inc., Germantown, WI, USA), and total RNA was isolated using an RNeasy Plant Mini Kit (Qiagen Inc., Hilden, Germany) according to the manufacturer’s manual. The purity and concentration of total RNA was measured using a Nanodrop spectrophotometer (Nanodrop ND-1000, Waltham, MA, USA). Total RNA was dissolved in DEPC-treated water, incubated at 65 °C for 10 min, and then transferred to ice. Then, 5 μg of total RNA was used to synthesize first-strand cDNA using an RNA to cDNA EcoDry Premix (Double Primed) (TAKARA, Kusatsu, Gunma, Japan). cDNA synthesis was performed at 42 °C for 60 min, and the enzyme was inactivated at 72 °C for 10 min. After cDNA synthesis, quantitative PCR was performed using TOPreal qPCR 2xPreMIX (SYBR Green with high ROX) (Enzynomics, Daejeon, Republic of Korea) on a StepOnePlus thermocycler (Applied Biosystems, Waltham, MA, USA). Rice UBIQUITIN 5 (OsUBI05) was used as an internal control and relative gene expression was analyzed by the 2−∆Ct or 2−∆∆Ct method.
Rna to cdna ecodry premix double primed
Manufactured by Takara Bio
Sourced in Japan, United States
The RNA to cDNA EcoDry Premix (Double Primed) is a convenient, ready-to-use solution for the reverse transcription of RNA to cDNA. It contains all the necessary reagents, including reverse transcriptase and random primers, in a lyophilized format for easy handling and storage.
Automatically generated - may contain errors
Lab products found in correlation
Topreal qpcr 2x premix sybr green with high rox, by Enzynomics (1 mentions)
Steponeplus thermocycler, by Thermo Fisher Scientific (1 mentions)
Rneasy plant mini kit, by Qiagen (1 mentions)
Sybr green qpcr mix, by Thermo Fisher Scientific (1 mentions)
Facscanto 2 flow cytometry, by BD (1 mentions)
Abi prism 7900pcr instrument, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Erythrocyte splitting liquor, by Tiangen Biotech (1 mentions)
2 protocols using rna to cdna ecodry premix double primed
1
RNA Extraction and qPCR Profiling in Rice
2
Quantitative Analysis of Immune Cell Activation
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TRIzol, 2X SYBR-Green qPCR mix, RevertAid™ Premium First Strand cDNA Synthesis kit, Hank's liquid, penicillin-streptomycin double antibodies and PBS were purchased from Invitrogen; Thermo Fisher Scientific, Inc. (cat. nos. 15596018, 4309155, K1621, 88284, 15070063 and 10010049; Waltham, MA, USA); reverse transcription kit from Takara Biotechnology Co., Ltd. (RNA to cDNA EcoDry™ Premix Double Primed; cat. no. 639549; Beijing, China); quantitative PCR kit from Takara Bio, Inc. (Otsu, Japan); CD69 monoclonal antibody (mouse anti-human phycoerythrin marker), CD3 monoclonal antibody (mouse anti-human activating protein), CD4 monoclonal antibody (BD Horizon™ BV510 mouse anti-human) and PE-labeled mouse IgG1 CD69 isotype control from BD Biosciences (dilution, 1:400; cat. nos. 560738, 552852, 562971 and 557908; Franklin Lakes, NJ, USA); erythrocyte splitting liquor from Tiangen Biotech Co., Ltd. (cat. no. RT122; Beijing, China); ABI Prism 7900 PCR instrument from Applied Biosystems (Thermo Fisher Scientific, Inc., Foster City, CA, USA), BD FACSCanto™ II flow cytometry from BD Biosciences.
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