Total RNA was extracted with TRIzol (Life Technologies, Foster City, CA, USA), according to the manufacturer's instructions. RNA quantification was performed by Qubit Fluorometer (Life Technologies). Contaminant DNA was removed using deoxyribonuclease 1 (DNase I Amplification Grade; Life Technologies). DNase-treated RNA was reverse transcribed by using a High Capacity RNA-to-cDNA Kit (Life Technologies), according to the manufacturer's instructions. Resulting cDNAs (30 ng per sample) were amplified through an ABI PRISM 7900HT Fast Real-Time PCR System (Life Technologies), as previously described (26 (link)). Single-gene specific assays were performed through real-time PCR by using Fast SYBR Green Master Mix (Life Technologies) according to the manufacturer's instruction. To allow statistical analysis, PCRs were performed in three independent biological replicates. The list of the primer pairs used for comparison analysis is reported below.
Deoxyribonuclease 1 dnase 1 amplification grade
Manufactured by Thermo Fisher Scientific
Deoxyribonuclease 1 (DNase I Amplification Grade) is an enzyme that catalyzes the hydrolytic cleavage of DNA. It is used to degrade DNA in RNA samples, preventing DNA contamination in downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Fast sybr green master mix, by Thermo Fisher Scientific (2 mentions)
Abi prism 7900ht fast real time pcr system, by Thermo Fisher Scientific (2 mentions)
High capacity rna to cdna kit, by Thermo Fisher Scientific (2 mentions)
Qubit fluorometer, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
2 protocols using deoxyribonuclease 1 dnase 1 amplification grade
1
Comprehensive RNA Extraction and Analysis
2
Comprehensive RNA Expression Analysis
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Total RNA was extracted with TRIzol (Life Technologies, Foster City, CA), according to the manufacturer's instructions. RNA quantification was performed by Qubit Fluorometer (Life Technologies). DNA contamination was removed using deoxyribonuclease 1 (DNase I Amplification Grade; Life Technologies). DNase-treated RNA was reverse transcribed by using High Capacity RNA-to-cDNA Kit (Life Technologies) according to manufacturer's instructions. Resulting cDNAs (200 ng per sample-loading port) were loaded into custom TLDAs, format 96a (Life Technologies), and amplified through a standard thermal cycling profile on an ABI PRISM 7900HT Fast Real-Time PCR System (Life Technologies). Single-gene specific assays were performed through real-time PCR by using Fast SYBR Green Master Mix (Life Technologies) according to manufacturer's instruction. To allow statistical analysis, PCRs were performed in three independent biological replicates. Primer sequences are available upon request.
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