Transgene Detection Primer Set for Real Time (Mouse)

Our Transgene Detection Primer Set for Real Time (Mouse) includes sets of primers designed to amplify sequences corresponding to GFP and lacZ, respectively—two markers commonly inserted into transgenic mice. These primer sets enable screening of transgenic mice by qPCR/real-time PCR.

Reference primer-1 and Reference primer-2 detect the mouse Ywhaz and Raver2 genes, respectively, and are included with the kit as references. They enable comparisons of relative transgene quantities between samples.

Cat. #	Product	Size	Price	License	Quantity	Details
3788	Transgene Detection Primer Set for Real Time (Mouse)	100 Rxns	USD $271.00
This kit is a primer set for real-time PCR to detect GFP (EGFP, AcGFP1) and lacZ, which are gene markers in transgenic mice. It is possible to screen transgenic mice by detecting these marker genes with real-time PCR. The primer sets (Ywhaz, Raver2) for detecting genes on mouse genomic DNA are also included as a reference. Therefore, it is possible to make a comparison of the transgene contents between samples by relative quantification. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Determination of homozygous/heterozygous genotypes using the Transgene Detection Primer Set for Real Time (Mouse) This experiment demonstrates how the Transgene Detection Primer Set for Real Time (Mouse) and quantitative PCR of genomic DNA from mouse tails can be used to accurately determine the homozygous and heterozygous genotypes of transgenic mice Determination of homozygous/heterozygous genotypes using the Transgene Detection Primer Set for Real Time (Mouse) This experiment demonstrates how the Transgene Detection Primer Set for Real Time (Mouse) and quantitative PCR of genomic DNA from mouse tails can be used to accurately determine the homozygous and heterozygous genotypes of transgenic mice. Five different mice representing three different genotypes for the EGFP and lacZ transgenes were used for the experiment. Tg#1 and Tg#3 were homozygous for the transgenes Tg#2 and Tg#4 were heterozygous wild-type (Wt) mouse had neither transgene* The mouse tails were kindly provided by Drs. Ikenaka and Takebayashi, National Institutes of Natural Sciences, Japan. Procedure* Genomic DNA was prepared from mouse tail samples using the FastPure DNA Kit. An aliquot (1 ul) of each DNA sample was then used as a template for real-time PCR using the primers included in the Transgene Detection Primer Set and Premix Ex Taq (Perfect Real Time) with a green intercalating dye. The transgene amplifications were quantified relative to the amplification signals produced from two endogenous reference genes. Results Relative quantification analysis using the ΔΔCt method demonstrated that samples Tg#1 and Tg#3 contain twice the EGFP and lacZ gene copy numbers contained in samples Tg#2 and Tg#4 (see figure). These results are consistent with actual genotypes of each sample. Neither transgene was detected in the Wt sample. Figure 1. Quantitative analysis of homozygous and heterozygous transgenic mice. Back 3788: Transgene Detection Primer Set for Real Time (Mouse)

More Information

Applications

Detection and quantification of mouse genome transgenes

Components

Sufficient for 100 reactions with each primer when using TB Green Premix Ex Taq II with 25 µl reaction volumes.

Product	Conc.	Amount	Size
GFP primer-1¹	2 µM each	500 µl	127 bp
GFP primer-2²	2 µM each	500 µl	162 bp
lacZ primer-1³	2 µM each	500 µl	96 bp
lacZ primer-2³	2 µM each	500 µl	141 bp
Reference primer-1⁴	2 µM each	500 µl	115 bp
Reference primer-2⁵	2 µM each	500 µl	89 bp

Notes:

¹GFP primer-1 is designed for detection of EGFP and AcGFP1 sequences.

²GFP primer-2 is designed for detection of EGFP sequence. It cannot be used for AcGFP1 detection.

³lacZ primer-1 and -2 are designed for lacZ (beta-galactosidase) gene detection and target different gene sequences.

⁴Reference primer-1 is designed for a part of the Ywhaz gene on mouse chromosome 15.

⁵Reference primer-2 is designed for a part of the Raver2 gene on mouse chromosome 4.

Storage

–20°C

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

Takara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2024 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.