Principle

Many mouse genotyping methods are based on the PCR amplification of genes of interest from tail biopsies or ear punches. The NucleoType Mouse PCR kit is designed for rapid mouse typing experiments and covers these standard sample materials including even blood and animal friendly hair samples. Hair samples can be easily collected causing minimal discomfort to the animal and do not require anesthesia. Thus, the NucleoType Mouse PCR kit enables animal friendly and time saving mouse genotyping.

A simple sample preparation within one tube is combined with a fast PCR. Mouse samples (tail clipping, ear punch, blood, or fur hair) are subjected to a quick lysis at room temperature and Proteinase K inactivation at 98 °C within 5 minutes. For optimal results an aliquot from the lysate is used as template for PCR typing.

The NucleoType HotStart PCR Master Mix (2x) enables a fast PCR (30–90 min) and contains a red dye for direct loading onto agarose gels. In addition the NucleoType Mouse PCR kit offers an alternative time-saving direct PCR protocol allowing PCR directly from unpurified and undiluted samples

 

Features

Technology	Simple sample preparation suitable for hot start PCR (optional direct PCR)
Format	Buffer and PCR mix (for 20–100 µL lysate and 10 µL PCR)
Sample material	Mouse tail clipping (1 mm), mouse ear punch (Ø 1 mm), mouse blood (1 μL), mouse hairs (approx. 3–30)
PCR	NucleoType HotStart PCR Master Mix for fast PCR genotyping (included)*
Amplicon size	Recommended for up to 1 kb
Preparation time	Lysis 2 min (18–25 °C); Enzyme inactivation 3 min (98 ° C); PCR cycling 30–90 min (cycler and target size dependent)
Stability	12 months at 4 °C or -20 °C

*For amplicons < 1 kb an extension time of approx. 15 sec is recommended.

 

Applications**

• Simple sample preparation in one single tube combined with a fast HotStart PCR
• Direct PCR for PCR analysis directly from unpurified and undiluted samples

 

Application data

Superior PCR results displaying strong bands within 30 minutes

Mouse EDTA-blood (male / female) purchased from Charles River Laboratories was used and 1 μL was processed according to manufacturer’s recommendations. Duplex PCR was performed for mouse genotyping (female ♀ one band / male ♂ two bands). PCR analysis of hair and ear samples was performed by amplification of a 466 bp and 273 bp fragment. Tail and blood samples were analyzed with primers for amplification of a 545 bp and 402 bp fragment. The Bioanalyzer results demonstrates competitive performance of the NucleoType Mouse PCR kit (MN) in comparison to all other tested kits. Fast and cost efficient sample preparation

 

 

Preparation times were compared between NucleoType Mouse PCR kit (MN) and several competitor kits. The unique lysis buffer of the NucleoType Mouse PCR kit enables template preparation within 5 minutes. An aliquot of this lysate is used as PCR template. Alternatively, direct PCR processing in higher PCR volumes is possible.

 

** Kits to be used for research purposes only