PDX models, short for patient-derived tumor xenografts (PDX), are xenografts created by implanting patient-derived tumor tissue and primary cells into immunodeficient mice. They are valuable tools for studying cancer mechanisms and testing drugs. However, tumor tissue fidelity can decrease with increasing passage number. Mouse cell infiltration not only affects model stability but can also lead to biased experimental data.
The StarLighter PDX Mouse Cell Infiltration Status Detection Kit accurately measures the concentration of human and mouse nucleic acids and assesses mouse cell infiltration status. The kit provides all necessary reagents for probe-based qPCR nucleic acid quantification. By generating standard curves for human and mouse nucleic acids, the absolute quantification method is used to calculate the human and mouse nucleic acid copy number concentrations in the test sample using these two standard curves. This effectively assesses mouse cell infiltration and ensures model fidelity.
| Product Introduction PDX models, short for patient-derived tumor xenografts (PDX), are xenografts created by implanting patient-derived tumor tissue and primary cells into immunodeficient mice. They are valuable tools for studying cancer mechanisms and testing drugs. However, tumor tissue fidelity can decrease with increasing passage number. Mouse cell infiltration not only affects model stability but can also lead to biased experimental data. The StarLighter PDX Mouse Cell Infiltration Status Detection Kit accurately measures the concentration of human and mouse nucleic acids and assesses mouse cell infiltration status. The kit provides all necessary reagents for probe-based qPCR nucleic acid quantification. By generating standard curves for human and mouse nucleic acids, the absolute quantification method is used to calculate the human and mouse nucleic acid copy number concentrations in the test sample using these two standard curves. This effectively assesses mouse cell infiltration and ensures model fidelity. Precautions
 Product Application The StarLighter PDX Mouse Cell Infiltration Status Detection Kit is a quantitative assay for human and mouse nucleic acids. Theoretically, this kit is compatible with human-to-mouse nucleic acid mixtures of varying ratios, with concentrations > 1 ng/µL. The lowest value of the standard curve for this kit is 10 copies/µL, enabling accurate quantification of mixtures with as little as 0.5% human nucleic acid. Product components  Storage and transportation conditions Store at -25 ~ -15℃ away from light, transport at < 0℃. Important parameters Standard dilution This kit provides a mixed human/mouse template (S1, 0.25 × 108 copies/μL) (hereinafter referred to as S1). The standard template is 0.25 × 108 copies/μL. S1 needs to be diluted 10-fold using the dedicated DNA diluent component provided in the kit to obtain S2. Continuous gradient dilution using the same method can obtain S2 to S6 (0.25 × 107 to 0.25 × 103 copies/μL). The diluted S2 to S6 can be stored at 2 to 8°C for up to 8 hours. If the storage time exceeds 8 hours, please dilute again. Sample quality Please use a qualified extraction kit to extract DNA from PDX model mouse tissues. Variations in extraction purity between different kits may cause fluctuations in qPCR Ct values. Use ultrapure water or Low TE for sample eluent. The maximum sample loading for this kit should not exceed 50 μg, and the maximum sample loading volume should not exceed 1/2 of the total volume. Accurate pipetting qPCR is a very sensitive technique, and this kit requires dilution of the standard, so the reliability of the results is highly dependent on accurate pipetting. To ensure accurate test results, please pay attention to the following:
Contamination and no-template controls Always adhere to good laboratory practices to avoid contamination of reagents, consumables, and equipment in the work area with samples, standards, or amplification products. It is recommended to include no-template controls (NTCs) in each assay to detect contamination introduced during reaction setup. The Cq/Ct value of the NTC should be at least 3 Cq/Ct higher than that of the diluted S6. During operation, to reduce contamination, ensure that standards are added to the reaction wells from low concentration to high concentration (ie, from S6 to S1). Replicate data, data reliability, throughput, and cost per sample qPCR is an extremely sensitive measurement technique susceptible to variability from multiple sources. It is recommended that triplicates be used for standards S1–S6, test samples, and negative controls. To increase throughput and reduce cost per sample, the number of replicates can be reduced to two. When choosing the optimal strategy for your workflow and throughput requirements, keep in mind that the reliability of your data is directly proportional to the number of replicates.
① S6 is < 3.1 cycles compared with S5, and the NTC is less than 3 Cq/Ct values after S6. ② If the amplification efficiency exceeds 100%, there may be contamination. ③ The baseline setting may delay the Cq/Ct value of S1 and affect the amplification efficiency. Manual adjustment of the baseline is required. ④ Inaccurate pipetting operation.
① Inaccurate pipetting operation. ② Improper operation of the instrument or improper storage of reagents.
① If the ΔCq/Ct between DNA S5 and S6 is < 3.1, there may be contamination. ② If the ΔCq/Ct between DNA S1 and S2 is < 3.1, there may be a problem with background subtraction and the baseline needs to be manually adjusted. ③ ΔCq/Ct > 3.6, poor amplification efficiency. Make sure all components are thoroughly mixed before use and confirm that all components are added in the correct volume. Add and use the correct amplification procedure. ④ Severe exposure of the reagent to light will reduce the total fluorescence and may cause a delay in the Cq/Ct value, resulting in a ΔCq/Ct value > 3.6. ⑤ Inaccurate pipetting operation and inaccurate standard dilution gradient.
① Inaccurate pipetting operation. ② Make sure all reagents are thoroughly mixed before use. ③ There is an instrument-related problem and you need to ensure that the correct reference dye is used.
① The sample condition results in, for example, very low nucleic acid content, below the detection limit. ② The sample input may be too high or too low.
① The sample to be tested is not pure enough and contains a large number of components that inhibit amplification. ② The sample concentration is too low. |
