Genotyping of miR-146a Knockout Mice

Mouse genotypes were confirmed by PCR^{42 (link),43 (link)}. Genomic DNA was isolated from tail snips using a Maxwell tissue DNA kit (Cat# AS1030, Promega, Madison, WI). miR-146a genotyping was performed using the following primers: 5′-GCTTATGAACTT GCCTATCTGTG-3′ and 5′-CAGCAGTTCCACGCTTCA-3′. PCR of wild-type and disrupted alleles generated amplicons of 350 bp and 159 bp, respectively. A representative gel of miR-146a+/+ and −/− amplicons is shown in the Fig. 1C ^{42 (link),43 (link)}.

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Javidan A., Jiang W., Okuyama M., Thiagarajan D., Yang L., Moorleghen J.J., Muniappan L, & Subramanian V. (2019). miR-146a Deficiency Accelerates Hepatic Inflammation Without Influencing Diet-induced Obesity in Mice. Scientific Reports, 9, 12626.

Publication 2019

Maxwell tissue DNA kit

Alleles Genomic Genotypes Mouse Primers Promega Tail Tissue

Corresponding Organization : University of Kentucky

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Variable analysis

independent variables

MiR-146a genotype (wild-type and disrupted alleles)

dependent variables

PCR amplicon size (350 bp for wild-type allele, 159 bp for disrupted allele)

control variables

Genomic DNA isolated from mouse tail snips using a Maxwell tissue DNA kit

controls

Positive control: miR-146a+/+ amplicon
Negative control: miR-146a-/- amplicon

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