RNA samples were collected 72 hr following siRNA transfection using the Quick-RNA Miniprep Kit (Zymo Research). Libraries were prepared by the VARI Genomics Core from 500 ng of total RNA using the KAPA mRNA HyperPrep kit (v4.17) (Kapa Biosystems). RNA was sheared to 300–400 bp. Prior to PCR amplification, cDNA fragments were ligated to IDT for Illumina unique dual adapters (IDT DNA Inc). Quality and quantity of the finished libraries were assessed using a combination of Agilent DNA High Sensitivity chip (Agilent Technologies), QuantiFluor® dsDNA System (Promega), and Kapa Illumina Library Quantification qPCR assays (Kapa Biosystems). Individually indexed libraries were pooled and 100 bp, single end sequencing was performed on an Illumina NovaSeq6000 sequencer using an SP, 100 cycle sequencing kit (Illumina) and each library was sequenced to an average raw depth of 35M reads. Base calling was done by Illumina RTA3 and output of NCS was demultiplexed and converted to FastQ format with Illumina Bcl2fastq v1.9.0.
Unique dual adapters
Manufactured by Illumina
Illumina unique dual adapters are specialized laboratory equipment designed to facilitate the preparation of DNA samples for sequencing analysis. These adapters are engineered to enable the attachment of unique molecular identifiers (UMIs) to DNA fragments, which is a crucial step in the sample preparation process. The core function of these adapters is to provide a mechanism for introducing UMIs into the DNA samples, allowing for the identification and tracking of individual DNA molecules during the sequencing workflow.
Automatically generated - may contain errors
Lab products found in correlation
Bcl2fastq v1, by Illumina (2 mentions)
Agilent dna high sensitivity chip, by Agilent Technologies (2 mentions)
Kapa mrna hyperprep kit v4.17, by Roche (2 mentions)
Novaseq 6000 sequencer, by Illumina (2 mentions)
Quantifluor dsdna system, by Promega (2 mentions)
Quick rna miniprep kit, by Zymo Research (2 mentions)
2 protocols using unique dual adapters
1
RNA-Seq Library Preparation and Sequencing
2
RNA-Seq Library Preparation and Sequencing
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RNA samples were collected 72 hr following siRNA transfection using the Quick-RNA Miniprep Kit (Zymo Research). Libraries were prepared by the VARI Genomics Core from 500 ng of total RNA using the KAPA mRNA HyperPrep kit (v4.17) (Kapa Biosystems). RNA was sheared to 300–400 bp. Prior to PCR amplification, cDNA fragments were ligated to IDT for Illumina unique dual adapters (IDT DNA Inc). Quality and quantity of the finished libraries were assessed using a combination of Agilent DNA High Sensitivity chip (Agilent Technologies), QuantiFluor® dsDNA System (Promega), and Kapa Illumina Library Quantification qPCR assays (Kapa Biosystems). Individually indexed libraries were pooled and 100 bp, single end sequencing was performed on an Illumina NovaSeq6000 sequencer using an SP, 100 cycle sequencing kit (Illumina) and each library was sequenced to an average raw depth of 35M reads. Base calling was done by Illumina RTA3 and output of NCS was demultiplexed and converted to FastQ format with Illumina Bcl2fastq v1.9.0.
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