Genotyping Mouse Ear Snips by PCR

DNA samples were prepared from mouse ear snips using NaOH extraction, and analyzed by PCR. PCR was performed using Quick-Load Taq 2× Master Mix (New England Biolabs). PCR conditions: denaturation at 94°C for 5 min followed by 30× 94°C for 30 s, 58°C for 30 s, 68°C for 45 s, 68°C for 5 min. Primers specific for Cre allele, internal control, floxed NR1 alleles were according to reference [25] (link). PCR products were analyzed by electrophoresis on 2% agarose with GelRed (Biotium).

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Pozzi L., Dorocic I.P., Wang X., Carlén M, & Meletis K. (2014). Mice Lacking NMDA Receptors in Parvalbumin Neurons Display Normal Depression-Related Behavior and Response to Antidepressant Action of NMDAR Antagonists. PLoS ONE, 9(1), e83879.

Publication 2014

Quick-Load Taq 2× Master Mix GelRed

Agarose Alleles Electrophoresis Mouse Primers

Corresponding Organization :

Other organizations : Karolinska Institutet

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Variable analysis

independent variables

NaOH extraction method for preparing DNA samples from mouse ear snips

dependent variables

PCR amplification of Cre allele, internal control, and floxed NR1 alleles
Electrophoresis analysis of PCR products on 2% agarose gel with GelRed

control variables

Quick-Load Taq 2× Master Mix (New England Biolabs) for PCR
PCR conditions: denaturation at 94°C for 5 min followed by 30× 94°C for 30 s, 58°C for 30 s, 68°C for 45 s, 68°C for 5 min
Primers specific for Cre allele, internal control, and floxed NR1 alleles according to reference [25]

controls

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

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