nucleic acid sequencing protocol
Standard Illumina clustering and sequencing protocol for Illumina GAIIx sequencer were used.
Mili and Miwi2 RNPs were immunoprecipitated as described in Reuter, M. et al. (2009). Small RNA libraries were generated with the protocol described in Hafner M. et al. (2008) but using adaptors suitable for sequencing on the Ilumina platform.
MiliDAH and Miwi2DAH mice were crossed with Oct4/GFP trangenic mice to label germ cells with eGFP. To isolate E16.5 and postnatal day 7 germ cells, single cell suspensions of testis were obtained by two step enzymatic digestion and GFP-positive FACS-sorted.
high throughput sequence alignment protocol
The reads were strippped of adapters and linkers, and low quality regions were masked. Only reads between length 24 and 30 were kept and aligned to mm9 genome using Bowtie.
nucleic acid library construction protocol
Small RNA libraries were generated with the protocol described in Hafner M. et al. (2008) but using adaptors suitable for sequencing on the Ilumina platform.