Total RNA was purified from cultured cells using Trizol (ThermoFisher Scientific) followed by treatment with Turbo™ DNase I (ThermoFisher Scientific). Enrichment of polyadenylated RNA [poly(A) RNA] was achieved through two rounds of selection with Oligo-(dT)25 Dynabeads (ThermoFisher Scientific) for LC-MS, dot blot or acRIP-seq, or two rounds of poly(A)Purist MAG (ThermoFisher Scientific) for ImmunoNorthern blot, according to the manufacturer’s instructions. Poly(A) RNA precipitations were carried out using 0.3M sodium acetate [pH 5.5], 15 μg/mL linear acrylamide (carrier) and 2.5× ethanol. Purification was estimated through Bioanalyzer picoRNA chips (Agilent Technologies) and by RT-qPCR using specific primers for 18S rRNA and GAPDH (see Table S7 for primer description). Briefly, RNA was reverse transcribed (RT) with random hexamers using the Superscript III system (ThermoFisher Scientific) according to the manufacturer’s suggestions, followed by qPCR using LightCycler 480 SYBR Green I Master Mix (Roche, Basil, Switzerland) in a LightCycler 96 Instrument (Roche).
Bioanalyzer picorna chips
Manufactured by Agilent Technologies
The Bioanalyzer picoRNA chips are a lab equipment product designed to analyze and quantify small RNA samples, such as microRNA (miRNA) and small interfering RNA (siRNA). The chips provide a rapid and sensitive method for the assessment of RNA integrity and concentration, enabling researchers to obtain reliable data for their RNA-based studies.
Automatically generated - may contain errors
Lab products found in correlation
Trizol, by Thermo Fisher Scientific (3 mentions)
Dynabeads oligo dt 25, by Thermo Fisher Scientific (1 mentions)
Superscript 3 system, by Thermo Fisher Scientific (1 mentions)
Turbo dnase 1, by Thermo Fisher Scientific (1 mentions)
Lightcycler 96 instrument, by Roche (1 mentions)
Lightcycler 480 sybr green 1 master mix, by Roche (1 mentions)
Hiseq 4000 platform, by Illumina (1 mentions)
Quantseq 3 mrna seq library prep kit fwd, by Lexogen (1 mentions)
2 protocols using bioanalyzer picorna chips
1
Enrichment of Polyadenylated RNA for Downstream Analysis
2
Total RNA Isolation and mRNA-Seq Library Prep
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was isolated from 50 OD600’s of cells as previously described [42 (link)] and libraries prepared as previously described [43 ]. In brief, cell pellets were lyophilized overnight and then RNA was isolated using TRIzol (Invitrogen) as previously described [42 (link)] and DNase treated as previously described [44 (link)]. 0.5 μg RNA was then prepared for sequencing using the QuantSeq 3‘-mRNA-Seq Library Prep Kit FWD (Lexogen) according to the manufacturer’s instructions. Input RNA quality and mRNA purity were verified by Bioanalyzer Pico RNA chips (Agilent). Libraries were sequenced on the HiSeq 4000 platform (Illumina).
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!