B. Differential PCR analysis was used to confirn the sperm-specific expression of F53G12.6. PCR products were amplified from cDNA libraries derived either from
fem-1(
hcl7ts) hermaphrodites which produce only oocytes, or from
fem-3(
q23ts) hermaphrodites which produce only sperm. The PCR reactions were multiplexed for three possible products: (i) a 169 bp product from F53G12.6 cDNA, (ii) a 526 bp product from F25H8.1, a somatic gene included as a non-germline-specific control, and (iii) a 714 bp product from
spe-12, included as a sperm-specific control. Both F53G12.6 and
spe-12 products appear sperm specific by their appearance only in the
fem-3 library, whereas the product from the somatic gene F25H8.1 was found in both libraries as expected. (PCR primers are given in Additional file 1).