Frozen spinal cord tissues were submerged in Trizol and homogenized in a Dounce homogenizer. Total RNA was extracted from the aqueous phase with the Quick-RNA Plus kit (Zymo, Irvine, CA) according to manufacturer instructions with on-column DNase I digestion. RNA quantity was measured by the Qubit RNA BR Assay Kit (ThermoScientific, Waltham, MA), and RNA integrity was assessed by the Bioanalyzer RNA Nano Eukaryote kit on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA).
Quick rna plus kit
Manufactured by Zymo Research
The Quick-RNA Plus Kit is a nucleic acid extraction system designed to efficiently isolate high-quality total RNA from a variety of sample types, including cells, tissues, and microorganisms. The kit utilizes a fast, spin-column-based method to ensure rapid and reliable RNA purification.
Automatically generated - may contain errors
Lab products found in correlation
Iscript cdna synthesis kit, by Bio-Rad (2 mentions)
Q5 polymerase, by New England Biolabs (2 mentions)
Cfx96 real time pcr detection system, by New England Biolabs (2 mentions)
Sybr green, by Lonza (2 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Qubit rna br assay kit, by Thermo Fisher Scientific (1 mentions)
Rna clean concentrator 5 kit, by Zymo Research (1 mentions)
Dynabeads myone streptavidin c1, by Thermo Fisher Scientific (1 mentions)
Quantus fluorometer, by Promega (1 mentions)
Spectrum plant total rna kit, by Merck Group (1 mentions)
Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
5 protocols using quick rna plus kit
1
Extraction and Quantification of Total RNA from Frozen Spinal Cord Tissue
2
Quantitative Real-Time PCR in HEK293T Cells
3
Testis RNA-seq Library Preparation
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We used approximately 100 pairs of testes from 3-5-day old mated w1118 males. The testes were dissected in 1X PBS and transferred into 200 µL RNAlater Solution. Tissue was pelleted by centrifuging at 5000g for 1 min at 4 °C. Supernatant was removed and 300 µL 1x DNA/RNA Shield was added before homogenization with an electric pestle. Homogenized tissue was digested with Proteinase K at 55 °C for at least 30 min. RNA was purified with the Zymo Quick-RNA Plus Kit (R1057).
Using up to 5 µg total RNA, ribosomal RNAs were removed suing iTools rRNA depletion Kit from Galen Laboratory Supplies (dp-P020-000007) and Thermo Fisher MyOne Streptavidin C1 Dynabeads (#65001). RNA Clean & Concentrator-5 kit from Zymo Research (R1015) was used to purify rRNA-depleted RNA. Starting with 1 ng-100 ng purified rRNA-depleted RNA, Illumina libraries were generated using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760).
Using up to 5 µg total RNA, ribosomal RNAs were removed suing iTools rRNA depletion Kit from Galen Laboratory Supplies (dp-P020-000007) and Thermo Fisher MyOne Streptavidin C1 Dynabeads (#65001). RNA Clean & Concentrator-5 kit from Zymo Research (R1015) was used to purify rRNA-depleted RNA. Starting with 1 ng-100 ng purified rRNA-depleted RNA, Illumina libraries were generated using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760).
4
Comprehensive RNA Extraction and Purification
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Total RNA was isolated with a combination of Trizol-based reagent RNA blue (Top-Bio, Vestec, Czech Republic), column-based system Quick RNA Plus Kit (Zymo Research, Irvine, CA, USA), and Wash buffer 1 from the column-based system Spectrum Plant Total RNA Kit (Sigma Aldrich, St. Louis, MO, USA). DNase treatment was carried out in a column. For confirming the expression of miRNAs using qPCR, small RNAs were captured from the total RNA using a Plant microRNA Purification Kit (Norgen, Thorold, ON, Canada) according to the kit protocol. RNA integrity was confirmed using 1.0% agarose gel electrophoresis, and the total RNA was quantified with a NanoDrop 1000 spectrophotometer (Thermo Scientific, Waltham, MA, USA) and a Quantus Fluorometer (Promega, Madison, WI, USA).
5
Quantitative Real-Time PCR in HEK293T Cells
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RNA was extracted from HEK293T cells using the Quick-RNA Plus Kit (Zymo Research). cDNA was synthesized using the iScript cDNA Synthesis Kit (Bio-Rad) and qPCR was performed on a Bio-Rad CFX96 Real-Time PCR Detection System using Q5 Polymerase (NEB) and SYBR Green (Lonza). Primers for qPCR are listed in Supplementary Table 10 .
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