Total RNA was extracted from 108 late log phase promastigotes using Trizol reagent as described by the manufacturer. RNA clean up was performed using RNeasy Plus mini kit (Qiagen, USA) according to manufacturer's instructions. The purified RNA was quantified using Nanodrop by estimating the absorbance at 260 and 280 nm. The quality and integrity of RNA was assessed on RNA 6000 Nano Assay Chips on Bioanalyzer 2100 (Agilent Technologies, USA) (Kulshrestha et al., 2014 (link)).
Rna 6000 nano assay chips on bioanalyzer 2100
Manufactured by Agilent Technologies
Sourced in United States
The RNA 6000 Nano Assay Chips on Bioanalyzer 2100 is a lab equipment product designed for the analysis of RNA samples. It provides quantitative and qualitative assessment of RNA samples. The product enables users to analyze the size, concentration, and integrity of RNA samples.
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2 protocols using rna 6000 nano assay chips on bioanalyzer 2100
1
Extracting High-Quality RNA from Promastigotes
2
RNA Isolation and Quality Evaluation
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Early log-phase promastigotes (1 × 108) of both K133WT and K133AS-R were used to isolate total RNA using TRIzol reagent according to the manufacturer’s instruction. Extracted RNA was cleaned up using a RNeasy Plus mini kit (Qiagen, Hilden, Germany). The absorbance of purified RNA was taken at 260 and 280 nm using a Nanodrop Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). The quality and integrity of RNA were assessed on an RNA 6000 Nano Assay Chips on Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA). RNA of good quality based on the 260/280 values (Nanodrop, Thermo Scientific, Waltham, MA, USA), rRNA 28S/18S ratios, and RNA integrity number (RIN) was used for further analysis [24 (link)].
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