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An amount of 0.9 ng of total RNA was added as competitor.
Then 100 ng of total RNA were reverse-transcribed with Super-Script II Reverse Transcriptase (Invitrogen).
cDNA was synthesized from 1000 ng of total RNA using iScriptTM cDNA Synthesis Kit (Biorad).
Real-time quantitative PCR (qPCR) was performed in a 15 μl reaction containing 200 800 ng of total RNA.
For qRT-PCR of bulk population samples, 10 ng of total RNA was used for each biological sample.
Briefly, 500 ng of total RNA was used to prepare labelled cRNA with overnight incubation according to the manufacturer's protocol.
The reverse transcription reaction was carried out using 10 ng of total small RNAs and the TaqMan advanced miRNA cDNA synthesis kit (Applied Biosystems).
500 ng of total RNA were amplified and labeled with Agilent Low Input Quick Amp Labeling Kit Agilent Technologiess, Palo Alto, CA, USA).
cDNA synthesis (100 ng of total RNA) was conducted using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, CA, USA).
5′ RACE (Invitrogen) was used to synthesize cDNAs from wildtype and albino axolotls using 250 ng of total RNA and gene-specific primers.
200 ng of total RNA was reverse transcribed using the High Capacity cDNA Synthesis kit (Life Technologies) and cDNA was subsequently diluted 1 10 in nuclease-free water.
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