Total RNA was extracted using both TRIzol (Invitrogen, Carlsbad, CA, United States) and the miRNeasy Mini Kit (QIAGEN, Valencia, CA, United States) according to the manufacturers’ protocols, with some modifications. Each sample was resuspended in 300 μl TRIzol reagent (Invitrogen). Resuspension was homogenized for 30 s using a Kontes Pellet Pestle Cordless Motor (Sigma-Aldrich, St. Louis, MO, United States) and placed on ice for 30 s, the steps being repeated 5 times for full homogenization. Then, an additional 700 μl TRIzol was added to the samples and incubated at room temperature for 5 min. 1-bromo-3-chloropropane (200 μl, Sigma-Aldrich) was then added to each sample, followed by incubation for 3 min. Next, the samples were centrifuged at 13,499 × g for 15 min at 4°C using a centrifuge (Smart R17 plus, Hanil Science, Incheon, South Korea). The supernatant (approximately 750 μl) of each sample was mixed with 750 μl of 100% isopropanol. The mixed samples were incubated at -20°C for 2 h. The columns of miRNeasy Mini Kit (QIAGEN) were additionally used to purify the small RNA-enriched RNA and clean RNA according to the manufacturer’s protocol.
Kontes pellet pestle cordless motor
Manufactured by Merck Group
Sourced in Macao
The Kontes Pellet Pestle Cordless Motor is a compact and portable motorized device used for grinding and homogenizing small samples. It features a rechargeable battery-powered motor that drives a serrated pestle to efficiently grind or mix samples in a variety of tube sizes.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
1 bromo 3 chloropropane, by Merck Group (1 mentions)
Mirneasy mini kit, by Qiagen (1 mentions)
Smart r17 plus, by Hanil (1 mentions)
Nebnext ultra 2 directional rna library prep kit, by New England Biolabs (1 mentions)
Nebnext poly a mrna magnetic isolation module, by New England Biolabs (1 mentions)
Qubit fluorometer, by Agilent Technologies (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Rlt buffer, by Qiagen (1 mentions)
2 protocols using kontes pellet pestle cordless motor
1
RNA Extraction Using TRIzol and miRNeasy
2
RNA-Seq protocol for larval transcriptomics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After 48 h treatment exposures surviving larvae were collected from three biological replicates per treatment, each biological replicate containing 15-30 surviving larvae. Frozen larvae were lysed in Buffer RLT (Qiagen), homogenized using a Kontes Pellet Pestle Cordless Motor (Sigma-Aldrich, St. Louis, Missouri), and total RNA extracted using the RNeasy mini kit (Qiagen). Total RNA concentration was quantified using a Qubit Fluorometer and RNA quality and integrity assessed using an Agilent 2100 Bioanalyzer. Only high-quality RNA samples with a RIN score > 7 on the Bioanalyzer were used for downstream library preparation.
mRNA was isolated from 500 ng of total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module, and the resulting mRNA prepared using the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs, Ipswich, Massachusetts). Final complementary DNA libraries were assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) to verify proper fragment sizes, and library concentrations were quantified using the Qubit Fluorometer. Libraries were multiplexed and sequenced as single-end 75 bp reads by the University of California Riverside genomics facility (Riverside, CA) in one lane of a NextSeq500 high-throughput sequencer (Illumina). Raw sequencing reads were submitted to the NCBI SRA database (Accession: PRJNA512294).
mRNA was isolated from 500 ng of total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module, and the resulting mRNA prepared using the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs, Ipswich, Massachusetts). Final complementary DNA libraries were assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) to verify proper fragment sizes, and library concentrations were quantified using the Qubit Fluorometer. Libraries were multiplexed and sequenced as single-end 75 bp reads by the University of California Riverside genomics facility (Riverside, CA) in one lane of a NextSeq500 high-throughput sequencer (Illumina). Raw sequencing reads were submitted to the NCBI SRA database (Accession: PRJNA512294).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!