title: GSM1263779: Degradome; Volvox carteri; RNA-Seq
library_construction_protocol: Gonidia and somatic cells were separated by Dounce homogenizer. Total RNAs were extracted from Volvox whole colonies, gonidia and somatic cells using TRIzol reagent (Invitrogen). Small RNAs were isolated from total RNA. Small RNAs were fractionated on a 15% PAGE gel. 18-28 nt small RNAs were purified and ligased to 3’ RNA linker-1 (IDT). 3’ ligation products were fractionated on 15% PAGE gel and 36~44 nt ligation products were purified and ligased to 5’ RNA linker. Ligation products were reverse transcribed by Supersript III (Invitrogen) and amplified by PCR. Poly(A)+ RNA were purified from total RNA by oligodT resin (Qiagen) and ligased to 5’ RNA linker. Ligation products were purified and reversed transcribed with RACER RT primer by Superscript III (Invitrogen). First strand cDNA were amplified by linker and RACER primers and digested by MmeI (NEB). Digestion products were purified and ligased to dsDNA linker. Ligation products were amplified by linker and dsDNA linker primers.
attributes.tag: GEO Accession
attributes.value: GSM1263779