Quantitative miRNA Expression Analysis

RNA extraction and real-time PCR were performed following the manufacturer’s protocols as previously reported (33 (link)–35 (link)). Briefly, total RNA (including small RNAs) were extracted from frozen tissues using mirVana miRNA Isolation Kits (Life Technologies). Reverse transcription (RT) was performed using TaqMan® MicroRNA Reverse Transcription Kits (Applied Biosystems, Foster City, CA), and the resulting cDNAs were quantitatively amplified for miR-34a, miR-30b and U6 using TaqMan® Universal Master Mix II, with UNG and TaqMan® MicroRNA specific primers for miR-34a (ID 000426), miR-30b (ID 000602) and U6 (ID 001973) on a 7900HT Fast Real-Time PCR System (Applied Biosystems, Foster City, CA). The relative miRNA expression fold change was quantified using the 2-ΔΔCT method with U6 as the endogenous control. Mean ± S.E.M was reported for N = 4 at each condition.

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Kiang J.G., Cannon G., Olson M.G., Smith J.T., Anderson M.N., Zhai M., Umali M.V., Ho K., Ho C., Cui W, & Xiao M. (2022). Female Mice are More Resistant to the Mixed-Field (67% Neutron + 33% Gamma) Radiation-Induced Injury in Bone Marrow and Small Intestine than Male Mice due to Sustained Increases in G-CSF and the Bcl-2/Bax Ratio and Lower miR-34a and MAPK Activation. Radiation research, 198(2), 120-133.

Publication 2022

mirVana miRNA Isolation Kits TaqMan® MicroRNA Reverse Transcription Kits TaqMan® Universal Master Mix II TaqMan® MicroRNA specific primers 7900HT Fast Real-Time PCR System

Cdnas Frozen Isolation Mirna Primers Real time pcr Reverse transcription Rnas Tissues

Corresponding Organization :

Other organizations : Uniformed Services University of the Health Sciences, American Association of Blood Banks, Johns Hopkins University, University of California, Los Angeles

Top 5 similar protocols

Variable analysis

independent variables

Extraction protocol (mirVana miRNA Isolation Kits)
Reverse transcription protocol (TaqMan® MicroRNA Reverse Transcription Kits)
Real-time PCR protocol (TaqMan® Universal Master Mix II, with UNG and TaqMan® MicroRNA specific primers)

dependent variables

Expression levels of miR-34a
Expression levels of miR-30b

control variables

Endogenous control (U6)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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