PMC:3091645 / 31841-32699 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/3091645","sourcedb":"PMC","sourceid":"3091645","source_url":"https://www.ncbi.nlm.nih.gov/pmc/3091645","text":"QPCR analysis\nThree μg of purified RNA was reverse transcribed with SuperScript II Reverse Transcriptase kit (Invitrogen) according to the manufacturer's protocol. Quantitative real-time PCR analysis was performed using ABI PRISM 7000 Sequence Detection System (Applied Biosystems) and SYBR Green (Applied Biosystems). Primers used are listed in Table 6. These primers were designed over exon-exon borders if possible. If not, a RT - control was added in the experiment to control for the absence of DNA contaminant. Normalization was done using the β-actin housekeeping gene. The temperature cycle used comprised 45 cycles at 95°C for 15 sec and 60°C for 1 min. A dissociation curve followed, this was comprised of 95°C for 15 sec, 60°C for 1 min and 95°C for 10 sec. Each sample was analysed in triplicate and data analysed using the Delta-Delta Ct method.","divisions":[{"label":"title","span":{"begin":0,"end":13}}],"tracks":[]}