PMC:7572125 / 52215-53312 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/7572125","sourcedb":"PMC","sourceid":"7572125","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7572125","text":"Quantitative PCR\nTotal RNA was extracted with TRIzol reagent (Life Technologies, cat# 15596026) and purified with RNeasy kit (Qiagen, cat# 74104) according to manufacturer’s instructions. RNA concentration and purity was determined (quality control data available on OSF project of each experiment - see Figure legends). Total RNA (1 µg) was reverse transcribed into cDNA using High Capacity cDNA Archive kit (Life Technologies, cat# 4368814) according to manufacturer’s instructions. Reactions consisted of cDNA (4.5 µl of 10X dilution), TaqMan Fast Advanced Mastermix (Life Technologies, cat# 4444964), and probes (TaqMan probes listed in Registered Report; Phelps et al., 2016). Reactions were performed in technical triplicate. Cycling conditions were: one cycle: 50°C for 2 min – one cycle: 95°C for 20 s – 40 cycles: [95°C for 1 min, 60°C for 20 s] using a StepOne Plus Real-Time PCR system (Applied Biosystems) and StepOne software (RRID:SCR_014281), version 2.3. Negative controls containing no cDNA template were included. Relative expression levels were determined using the ∆∆Ct method.","divisions":[{"label":"title","span":{"begin":0,"end":16}}],"tracks":[{"project":"2_test","denotations":[{"id":"33073769-26943900-27525653","span":{"begin":675,"end":679},"obj":"26943900"}],"attributes":[{"subj":"33073769-26943900-27525653","pred":"source","obj":"2_test"}]},{"project":"MyTest","denotations":[{"id":"33073769-26943900-27525653","span":{"begin":675,"end":679},"obj":"26943900"}],"namespaces":[{"prefix":"_base","uri":"https://www.uniprot.org/uniprot/testbase"},{"prefix":"UniProtKB","uri":"https://www.uniprot.org/uniprot/"},{"prefix":"uniprot","uri":"https://www.uniprot.org/uniprotkb/"}],"attributes":[{"subj":"33073769-26943900-27525653","pred":"source","obj":"MyTest"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#ecb193","default":true},{"id":"MyTest","color":"#9793ec"}]}]}}