1. Evaluation of ribonucleic acid amplification protocols for human oocyte transcriptome analysis 
Eleni Mantikou, Ph.D., Oskar Bruning, M.Sc., Sebastiaan Mastenbroek, Ph.D., Sjoerd Repping, Ph.D., Timo Markus Breit, Ph.D., Mark de Jong, Ph.D. 
Fertility and Sterility 
Volume 105, Issue 2, Pages e4 (February 2016)
DOI: /j.fertnstert
Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Amplification quality. Examples of the Bioanalyzer electrophoretic profiles of amplified products (single oocyte: black line; positive control: blue line; NTC: green line) from four amplification protocols.
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3. Figure 2
Quality and quantity assessment of amplification products. (A) Examples of the Bioanalyzer electrophoretic' profiles of cDNA obtained using NuGEN WT-Ovation One direct and aRNA obtained using Arcturus RiboAmp HS Plus (single oocyte: black line; pooled-and-split oocyte: grey line; positive control: blue line; NTC: green line). (B) Complementary DNA and aRNA amplification yields obtained with the two protocols.
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4. Figure 3
Validation of the protocols' performance on gene-expression profiling. (A) Principal component analysis of the unnormalized log2 ratios (test/reference). (B) Spearman correlations showing the similarity of the unnormalized microarray data from single and pooled-and-split samples obtained by the two protocols. (C) Mean number of present transcripts per protocol. (D) Number of differentially expressed genes per protocol.
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5. Supplemental Figure 1
Amplification protocols. Amplification process of NuGEN WT Ovation One-Direct (left) and Arcturus RiboAmp HS PLUS (right).
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6. Supplemental Figure 2
Experimental design for validating the two protocols. Pooled-and-split oocytes (NPS and RPS) were used to determine the technical variation introduced by the two protocols, and single oocytes (NS and RS) were used to determine the biological variation among single human oocytes.
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7. Supplemental Figure 3
Heatmap of the top 300 expressed genes. The genes that overlap within a protocol are colored, and the ones not overlapping are presented in grey.
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8. Supplemental Figure 4
GC content of the DEGs in the two protocols. Comparison of GC content for the differentially expressed genes in samples amplified by NuGEN WT Ovation One direct, Arcturus RiboAmp HS PLUS, and all samples together.
Fertility and Sterility  , e4DOI: ( /j.fertnstert )
Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions