Glomus Titanium Simple
Aliquots of about 100,000 sterile G. intraradices spores from in vitro culture were activated in 6ml of M medium containing 1.5.10-6 M apigenin for 24h at 30 C, 2% CO2. Spores were immediately frozen in liquid nitrogen. Total RNA was extracted using a RNeasy Plant Mini Kit (Qiagen) with RLT lysis buffer. RNA quality and integrity were checked prior to cDNA synthesis using the Bio-Rad Experion analyzer and Experion RNA StdSens analysis kit. Total RNA preparations (three biological replicates) were amplified using the SMART PCR cDNA Synthesis Kit (Clontech) according to the manufacturer's instructions. cDNAs were pooled after purification using QIAquick PCR purification Kit (Qiagen) and used for 454 pyrosequencing.