RNA extraction, lllumina sequencing and downstream data analyses were performed as described previously (29 (link), 20 (link)). RNA was extracted from each sample of 30 adult flies. There were three replicate samples for each 4-genotype x 3-hypoxia x 2-sex treatment conditions, with the exception of only two replicates from the female siI;Austria genotype at time point t = 0, resulting in a total of 71 RNA libraries. Analyses reported here are for 35 libraries from female samples. RNA extraction followed procedures described in (30 (link)) for mRNA purification, fragmentation, first and second strand cDNA synthesis, adapter ligation and PCR enrichment. Nucleic acid quantification at each stage was performed using a Qubit fluorimeter using Qubit reagents. Libraries for RNAseq were prepared from amplified cDNA fragments ranging from 334–500 bp using a Caliper Lab Chip XT apparatus with DNA 750 chips (Caliper Life Sciences, Hopkinton, Massachusetts USA). Quantification of transcript levels was performed using 50 bp single end sequence reads on an Illumina HiSeq 2000 instrument at the Brown University Genomics Core Facility.
Dna 750 chips
Manufactured by PerkinElmer
Sourced in United States
The DNA 750 chips are a type of laboratory equipment designed for genomic analysis. They are used to process and analyze DNA samples. The chips facilitate the detection and quantification of genetic information.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using dna 750 chips
1
RNA-seq Analysis of Hypoxia Responses
2
Transcriptomic Analysis of Hypoxia Response
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RNA extraction, Illumina sequencing and downstream data analyses were performed as described previously 20 , 29 . RNA was extracted from each sample of 30 adult flies. There were three replicate samples for each 4‐genotype × 3‐hypoxia x 2‐sex treatment conditions, with the exception of only two replicates from the female siI;Austria genotype at time point t = 0, resulting in a total of 71 RNA libraries. Analyses reported here are for 35 libraries from female samples. RNA extraction followed procedures described in 30 for mRNA purification, fragmentation, first and second strand cDNA synthesis, adapter ligation, and PCR enrichment. Nucleic acid quantification at each stage was performed using a Qubit fluorimeter using Qubit reagents. Libraries for RNAseq were prepared from amplified cDNA fragments ranging from 334–500 bp using a Caliper Lab Chip XT apparatus with DNA 750 chips (Caliper Life Sciences, Hopkinton, MA). Quantification of transcript levels was performed using 50 bp single end sequence reads on an Illumina HiSeq 2000 instrument at the Brown University Genomics Core Facility.
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