cell type: single primordial germ cell genotype: Prdm14 (+/+)
Extracted molecule
total RNA
Extraction protocol
C57BL/6 mice with the Prdm14 (+/-) genotype (Yamaji et al., Nature Genetics, 2008) were crossed. Pregnant mice were sacrificed at embryonic day (E) 7.25 - 7.5 and embryos at the early-to-mid allantois bud stage were isolated. The base of the allantois bud, where primordial germ cells (PGCs) were clustered, was dissected using a glass knife and cells were dissociated with trypsin as described (Kurimoto et al., Genes and Development, 2008). Single cells were randomly isolated manually using a glass capillary as described (Kurimoto et al., Nature Protocols, 2007) and directly subjected to cDNA amplification (Kurimoto et al., Nucleic Acids Research, 2006). cDNAs of PGCs were identified based on Prdm1 expression using targeted PCR as described (Kurimoto et al., Genes and Development, 2008). Genotypes of Prdm14 were identified by PCR as described (Yamaji et al., Nature Genetics, 2008). cDNAs of PGCs with Prdm14 (+/+), (+/-), and (-/-) genotypes were then subjected to microarray analysis using the Affymetrix GeneChip Mouse Genome 430 2.0 Array.
Label
biotin
Label protocol
not provided
Hybridization protocol
not provided
Scan protocol
not provided
Description
Gene expression data from a single primordial germ cell isolated using a glass capillary Prdm14 (+/+) PGC at early-to-late bud stage (E7.25-E7.5)
Data processing
Genome 430 2.0 Array. Microarray data was processed using dChip (Ver.1.3) with the PM/MM difference model. Expression level of Dppa3 (1424295_at) were investigated.
