RNA was isolated from the pooled whole worm samples in TRIzol reagent according to the manufacturer’s instructions (Life Technologies). For the male head samples, ten additional male worms per mouse per time point were dissected in vivoPHIXTM by cutting posterior to the ventral sucker and anterior to the testes. The heads were rinsed in 50% ethanol and pooled in TRIzol and the RNA extracted as for the whole worm samples. RNA quality was assessed using a Pico RNA kit for the BioAnalyzer (Agilent). Good quality RNA was extracted from all but one sample (male head day2_20:00_c). Total RNA was enriched for mRNA using poly(A) pulldown. The sequencing libraries were prepared using the NEB Ultra II RNA custom kit on an Agilent Bravo WS automation system. All samples had 14 cycles of PCR, which was set up using Kapa HiFi Hot start mix and Eurofins dual indexed tag barcodes on Agilent Bravo WS automation system. Paired-end 75-nucleotide RNA sequencing reads were produced from the pooled worm libraries across six lanes of an Illumina HiSeq2500 v4 sequencing platform. All sequencing data are available through ENA study accession number ERP108923 [97 ].
Pico rna kit for the bioanalyzer
Manufactured by Agilent Technologies
The Pico RNA kit for the BioAnalyzer is a laboratory equipment product designed to analyze small amounts of RNA samples. It provides a sensitive and accurate method for quantifying and assessing the quality of RNA samples.
Automatically generated - may contain errors
2 protocols using pico rna kit for the bioanalyzer
1
Transcriptome Analysis of Schistosoma Mansoni
2
Transcriptome Analysis of Whole Worms and Male Heads
Check if the same lab product or an alternative is used in the 5 most similar protocols
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RNA was isolated from the pooled whole worm samples in TRIzol reagent according to the manufacturer's instructions (Life Technologies). For the male head samples, ten additional male worms per mouse per time point were dissected in vivoPHIX TM by cutting posterior to the ventral sucker and anterior to the testes. The heads were rinsed in 50% ethanol and pooled in TRIzol and the RNA extracted as for the whole worm samples. RNA quality was assessed using a Pico RNA kit for the BioAnalyzer (Agilent). We were able to extract good quality RNA from all but one sample (male head day2_20:00_c). Total RNA was enriched for mRNA using poly(A) pulldown. The sequencing libraries were prepared using the NEB Ultra II RNA custom kit on an Agilent Bravo WS automation system. All samples had 14 cycles of PCR, which was set-up using Kapa HiFi Hot start mix and Eurofins dual indexed tag barcodes on Agilent Bravo WS automation system. RNA sequencing of the pooled worm libraries was performed on six lanes of the Illumina HiSeq2500 v4 75 Paired End sequencing platform. All sequencing data are available through ENA study accession number ERP108923.
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