PP-D050T Pichia Pastoris Host Cell DNA Residue Detection kit
The Pichia Pastoris Host Cell DNA Residue Detection Kit can be used for Quantitative analysis of DNA contaminants in recombinant protein expression, intermediate purification, and finished production from the host cell.
Introduction |
This kit adopts Taqman probe fluorescence qPCR method. The kit has the advantages of high specificity and sensitivity by using specific primers & probes, LOD can reach 1fg level.The preparation process of DNA Control is completely consistent with National Standard, therefore it has high purity and no protein and ion interference to ensure the accuracy of the sample quantitative detection.The kit provides DNA Dilution Buffer, which enables good replicate parallelism in a single experiment and good reproducibility between multiple experiments. |
| Kit Components | ||
DNA Amplification | ||
Components NO. | Components Name | Cat#/Size PP-D050T(50T) |
B1 | 2XqPCR Mix | 0.625mL |
B2 | Primer&Probe Mix | 100μL |
B3 | 2×1.5mL | |
B4 | DNA Control (10ng/μL) | 25μL |
B5 | RNase-Free H2O | 0.5mL |
B6 | 50X ROX Reference Dye(Optional) | 0.15ml |
Shipping and Storage | |
| 1 | All components are shipped on dry ice. |
| 2 | The kit should be stored at -20°C and has a shelf life of two years. B2 should be stored protected from light. |
Accuracy (Standard: ≦30%) | Intra Variation% 3.1-10 |
Inter Variation% 10-24 | |
Recovery | 69-127% |
| Limit of Quantitation | 10fg/μL |
Specificity | - |
Components | Volume(μL) |
2XqPCR Mix | 12.5 |
Primer&Probe Mix | 2 |
DNA template (control or sample) | 5 |
Add water | 5.5 |
Total Volume | 25 |
Mix solution = (number of reaction wells+4) * (12.5+2+5.5)μL (including the volume lost in the 4 wells).
The detection range of the standard curve mentioned above is suitable for most experiments and can be adjusted as needed, such as 3fg/ul-300pg/ul.
Standard Curve: R ²> 0.99; Amplification Efficiency: 90% ≤ Eff% ≤ 110%; Slope: -3.8~-3.1.
The recovery rate of spiked samples=(measured value of spiked samples - measured value of samples)/theoretical value of spiked samples * 100%, with a range of 50% -150%.
Negative Quality Control (NC): The reaction system contains only DNA Dilution Buffer, and the detection result should have a Ct value greater than the mean Ct value of the lowest concentration Ct on the standard curve.
No Template Control (NTC): Replace only the DNA template with DNA Dilution Buffer in the reaction system, and the Ct value obtained should be 'Undetermined' or ≥35.
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