Evaluation of ribonucleic acid amplification protocols for human oocyte transcriptome analysis
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ABSTRACT: Objective: To develop a reliable, reproducible, and sensitive method for investigating gene-expression profiles from individual human oocytes. Setting: University medical center and university microarray laboratory. Patient(s): Couples undergoing intracytoplasmic sperm injection treatment were asked to donate their immature oocytes for research, and written informed consent was obtained in all cases. Seventy-nine human oocytes were used in total. Intervention(s): None. Main Outcome Measure(s): Amplification efficiency and microarray profiles. Result(s): Two of the five protocols (WT-Ovation One-Direct and Arcturus RiboAMp HS Plus) resulted in sufficient yields and high success rates and were further validated for their performance in obtaining reliable, reproducible, and sensitive expression profiles from individual human oocytes. Evaluation of these two protocols demonstrated that they both displayed low technical variation and produced highly reproducible profiles (r R 0.95). One of them identified significantly more transcripts but also had a higher number of false discoveries. Conclusion(s): Two protocols generated ample amounts of mRNA for (quantitative) polymerase chain reaction, microarray, and sequencing techniques. Further validation using a design that discriminates between biological and technical variation showed that both protocols can be used for gene-expression profiling of individual human oocytes.
ORGANISM(S): Homo sapiens
PROVIDER: GSE48141 | GEO | 2015/12/31
SECONDARY ACCESSION(S): PRJNA209022
REPOSITORIES: GEO
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