Total RNA from left axillary mammary gland and liver samples were isolated by acid guanidinium isothiocyanate–phenol–chloroform extraction (TRIzol Reagent, Invitrogen, Rockville, MD, USA) (1 ml of solution per 100 mg of tissue) and further purified by the RNeasy Midi Cleanup Kit (Qiagen, Valencia, CA, USA) in accordance with the manufacturers’ protocols. Total RNA concentrations were determined by spectrophotometric evaluation of absorbance at 260 nm, and RNA quality and integrity were confirmed by 260/280 nm ratio (Nanodrop2000, UV-Vis Spectrophotometer, Thermo Scientific, Wilmington, DE, USA) and 1% agarose RNA gel electrophoresis respectively. The RNA quality of random samples was also tested on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA).
Rneasy midi cleanup kit
Manufactured by Qiagen
Sourced in United States
The RNeasy Midi Cleanup Kit is a laboratory product designed to purify and concentrate RNA samples. It utilizes a silica-membrane technology to efficiently capture and recover RNA molecules from various biological samples. The kit is intended to facilitate the cleanup and concentration of RNA for downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Nanodrop 2000 uv vis spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Quantstudio 3 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Lightcycler 480 sybr green 1 master, by Roche (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using rneasy midi cleanup kit
1
Comprehensive RNA Isolation from Mammary and Liver Tissues
2
Quantification of Pcdhg Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Whole brains were collected from mice at P21 and RNA isolated using TRIzol reagent (ThermoFisher) according to the manufacturer’s protocol. RNA was further purified using the RNeasy midi cleanup kit (Qiagen) according to the manufacturer’s protocol. Two μg of total RNA was used for cDNA synthesis using High-Capacity cDNA Reverse Transcription kit (ThermoFisher). The cDNA produced was diluted 1:2 for use with LightCycler 480 SYBR Green I Master (Roche) with the primers listed in the section above. The QuantStudio 3 Real Time PCR system (ThermoFisher) was used to carry out qPCR reactions. Relative Pcdhg abundance was calculated using the ΔΔCt method, normalized to GAPDH and control mice.
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