GSM1437523
GSM1437523: Argonaute IP; Volvox carteri; RIP-Seq
myc-tagged Argonaute was transformed into vegetative growing Volvox carteri, strain Vol6. Colonies were pelleted and sheared to dissociate the cells from the extracellular matrix. After centrifugation, cells were lysed in lysis buffer with additional sonication. The immunoprecipitation (IP) was performed using the cleared lysate with anti-myc antibodies. After washing of the IP, the RNA was recovered by performing first a Proteinase K digest followed by a phenol/chloroform extraction. The RNA was precipiated over night with Ethanol, recovered by centrifugation and solved in ultra-pure water. The Library was constructed according to Illumina's TruSeq small RNA cloning kit using the following adapters:  5′adapter 5′-GUUCAGAGUUCUACAGUCCGACGAUC-3′; 3′adapter Phospho-TGGAATTCTCGGGTGCCAAGG-Amino-C7. The index was added by PCR later in the protocol: Used index:  Sample 1=Index 12; 5' PCR primer 5'-AATGATACGGCGACCACCGAGATCTACACGTTCAGAGTTCTACAGTCCGA-3'; 3' PCR primer 5'-CAAGCAGAAGACGGCATACGAGATTACAAGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA-3'
GEO
GSM1437523
GEO Accession
GSM1437523