Description

[Product Specifications] PDX50 50T

[Product Description] Patient-Derived tumor Xenograft (PDX) models are established by directly transplanting patient tumor tissues into immunodeficient mice. While these models serve as valuable tools for cancer mechanism research and drug testing, the tumor tissue fidelity decreases with increasing passage numbers.

This kit uses dual-color fluorescent probe qPCR (FAM+HEX) to detect the proportion of mouse DNA in total DNA from PDX models, thereby inferring mouse cell infiltration ratio. The detection target is tissue DNA. The kit features the following characteristics: Stable: Closed-tube operation prevents cross-contamination.

1. Reliable: Samples tested alongside multiple internal standards on the same plate, reducing well-to-well interference.
2. Convenient: Simple operation, no electrophoresis required.
3. Quantitative: Quantitative detection using ΔCT values.

[Transportation and Storage Conditions] Shipped frozen at low temperature, store at -20°C, shelf life 1 year. After thawing, store at 4°C for up to 4 weeks.

[Kit Components] [Table with components]

Notes: (1) Biowing®PDX qPCR SuperMix contains primers and other PCR components; (2) Standards contain mouse DNA and human genomic DNA, with Standards 1, 2, and 3 corresponding to 25%, 50%, and 75% mouse DNA content respectively; (3) Before using the provided reagents, gently invert ten times to mix, avoid bubbles, centrifuge briefly and open carefully.

[Threshold Settings] Using Standard2 (50%) amplification curve, set threshold line to meet following CT value ranges: (1) Human DNA amplification curve CT value set to 25±1 (2) Mouse DNA amplification curve CT value set to 24±1

[Operating Procedures]

1. Sample Preparation Extract tissue DNA using commercial DNA extraction kit. DNA purity A260/A280≈1.8, recommended concentration 20-50ng/µL, load 2µL. Note: Excessively high or low sample concentrations affect amplification efficiency, interfering with experiments and increasing measurement error.
2. qPCR Reaction System and Conditions 20µL reaction system example

PCR instrument settings using SLAN-96S as example

[Standard Curve] Example

Note: Recommended to generate standard curve for each batch tested, using three positive standards with three technical replicates each. Plot scatter diagram with linear fitting trend line to obtain regression equation for calculating mouse cell infiltration ratio.

[Result Interpretation] Using standard curve linear regression equation: Y=-0.4061X+0.4089 as example Cell ratio calculation method in PDX model (mouse:human): Calculate mouse and human cell ratios in PDX samples based on standard curve. Note: Standard curve linear regression equation: Y=-0.4061X+0.4089 Correlation coefficient R2: 0.9998 X: ΔCt value (CT mouse – CT human) Y: log10 value of mouse:human mixture ratio in PDX model, i.e., Y=log10(mouse:human ratio) Mouse:human ratio = power(10,Y) Mouse DNA percentage of total DNA = power(10,Y)/(1+ power(10,Y)) Human DNA percentage of total DNA = 1 – Mouse DNA percentage of total DNA