RNA expression levels for GSK3β and APBB2 were obtained for the ROS/MAP from frozen sections of the dorsolateral prefrontal cortex that was manually dissected from postmortem brain tissue. Details of RNA extraction, processing and data quality control and normalization have been previously published (Lim et al. 2014 (link)). In brief, RNA was isolated using the RNeasy lipid tissue kit (Qiagen, Valencia, CA) and was reverse transcribed and biotin-UTP labeled using the llumina® TotalPrep™ RNA Amplification Kit from Ambion (Illumina, San Diego, CA). Expression signals were generated using the Beadstudio software suite (Illumina, San Diego, CA). Standard control and normalization methods were employed to account for technical variability due to differences in hybridization dates and stabilize the variance for the purpose of statistical analyses.
Llumina totalprep rna amplification kit
Manufactured by Thermo Fisher Scientific
The Illumina® TotalPrep™ RNA Amplification Kit is a laboratory product designed for the amplification of RNA samples. It provides a method for generating biotinylated, amplified, single-stranded complementary DNA (cDNA) from small amounts of total RNA for use in gene expression profiling.
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2 protocols using llumina totalprep rna amplification kit
1
Quantifying RNA Expression in Brain Tissue
2
Transcriptional Profile of Maternal Cells in Smoking
Check if the same lab product or an alternative is used in the 5 most similar protocols
Votavova et al. (2011 (link)) assessed the effect of smoking on maternal cells at the transcription level. In the study, they collected blood samples of 52 nonsmokers and 20 smokers whose ages ranged from 18 to 41. However, only 46 nonsmokers and 19 smokers were engaged for the following analysis. Sample information used in this study is given inSupplementary Table S1 . Expression levels of 24,526 transcripts in these samples were detected by the HumanRef-8 v3 Expression BeadChips (Illumina, San Diego, CA, United States). Based on the data (GSE27272), we aimed to screen age-associated mRNAs. Detailed experimental procedures were reported in the study (Votavova et al., 2011 (link)). In brief, RNA samples were extracted and purified from blood samples by using the LeukoLOCK™ Total RNA Isolation System (Ambion, Austin, TX, United States). Second, cRNA was synthesized and biotinylated by using the llumina TotalPrep RNA amplification kit (Ambion). The hybridization reaction of each cRNA sample was conducted on the beadchips and scanned by using the BeadArray Reader. Finally, the obtained raw data were processed and normalized by the quantile method in the Lumi package of R software (www.r-project.org ).
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Votavova et al. (2011 (link)) assessed the effect of smoking on maternal cells at the transcription level. In the study, they collected blood samples of 52 nonsmokers and 20 smokers whose ages ranged from 18 to 41. However, only 46 nonsmokers and 19 smokers were engaged for the following analysis. Sample information used in this study is given in
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