BlueGene Biotech Pichia Pastoris Host Cell DNA Residue Detection kit-50T

 PP-D050T Pichia pastoris HCD 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.

 

Specifications of Pichia Pastoris Host Cell DNA Residue Detection kit

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	DNA Dilution Buffer	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.

 

 

Quality Control on Pichia Pastoris Host Cell DNA Residue Detection kit

Accuracy (Standard: ≦30%)	Intra Variation% 3.1-10
	Inter Variation% 10-24
Recovery	69-127%
Limit of Quantitation	10fg/μL
Specificity	-

 

PCR Reaction System on Pichia Pastoris Host Cell DNA Residue Detection kit

 

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.

 

Criteria for Results

 

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.