RNA was purified using the ZR RNA MicroPrep kit (Zymo). Nugen RiboSpia Ovation Pico WTA System V2 was used for target amplification for RNA-seq. For microarrays we used the SCAMP method previously described (Brunskill et al., 2011c (link)). RNA-seq was carried out using 50b single end reads on the Illumina Hi-Seq 2500 machine according to Illumina protocols, with read depths > 40 million. For microarrays we examined a minimum of three biological replicates, and commonly four to six. Over one hundred microarrays in total were used. Exact replicate numbers are shown in heatmaps and at FaceBase.Org. For RNA-seq single samples were analyzed.
Pico wta system v2
Manufactured by Tecan
The Pico WTA System V2 is a fully automated, high-throughput library preparation solution for whole transcriptome amplification. It is designed to generate amplified cDNA libraries from small amounts of total RNA samples for use in next-generation sequencing applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pico wta system v2
1
Transcriptomic Profiling of Biological Samples
2
FACS-Isolated Melanoma RNA Amplification
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To further validate candidate gene expression profiles obtained from the RNAseq datasets, metastatic melanoma cells were FACS isolated as described above from independent experimental samples. The resultant RNA were in pg/μl range and this was amplified using the Nugen Ovation® Pico WTA System V2. RNA amplification was performed based on the protocol and instructions provided in the kit using the Ribo-SPIA technology.
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