RNA Isolation and RT-PCR Analysis

Total RNA was isolated using the RNeasy Mini Kit (Qiagen GmBH, Hilden, Germany). From each sample, 2 μg of RNA was converted into cDNA by oligo(dT)₁₈-primed reverse transcription using SuperScript II RT First-Strand kit (Invitrogen, Carlsbad, CA, USA) as described by the manufacturer. The cDNA was subject to semiquantitative PCR analysis using Accuprime Taq polymerase system (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s recommendations. Flt3 primers were used as previously described in Ref. (8 (link)).

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Nin D.S., Li F., Visvanathan S, & Khan M. (2015). Misfolded N-CoR is Linked to the Ectopic Reactivation of CD34/Flt3-Based Stem-Cell Phenotype in Promyelocytic and Monocytic Acute Myeloid Leukemia. Frontiers in Oncology, 5, 210.

Publication 2015

RNeasy Mini Kit SuperScript II RT First-Strand kit Accuprime Taq polymerase system

Cdna Flt3 Oligo Primers Reverse transcription Taq polymerase

Corresponding Organization :

Other organizations : National University of Singapore, AIMST University

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Variable analysis

independent variables

RNA isolation using RNeasy Mini Kit
Reverse transcription using SuperScript II RT First-Strand kit
Semiquantitative PCR analysis using Accuprime Taq polymerase system
Flt3 primers used as previously described in Ref. (8)

dependent variables

CDNA levels

control variables

Amount of RNA (2 μg) used for cDNA conversion
Oligo(dT)18-primed reverse transcription
Manufacturer's recommendations for SuperScript II RT First-Strand kit and Accuprime Taq polymerase system

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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