Cells were propagated in RPMI 1640 medium with 10% fetal calf serum, 100 ug/ml each streptomycin and penicillin, at 37C with 5% C02.
Extracted molecule
polyA RNA
Extraction protocol
Cell monolayers were disrupted with RLT+ lysis solution and total cellular RNA isolated with RNeasy kits, treated with DNaseI, followed by a second RNA isolation. PolyA RNA was enriched from total RNA using PolyATract mRNA isolation kit.
Description
Standard SAGE protocols were used to generate library, cloned into pZER0-1, and sequenced on an ABI 3700 system.
Data processing
Data were processed using Phred and Sequencher with manual editing then processed by eSAGE software.
