E.coli HCD Residue Detection Kits

 BlueGene Biotech

E. coli Host Cell DNA Residue Detection kit-50T

 

EC-D050T E.coli Host Cell DNA Residue Detection kit

 

The E.coli HCDA Residue Detection Kit can be used for Quantitative analysis of DNA residue in recombinant protein expressed products, purified intermediate and finished products from the host cell.

 

Specifications of E.coli 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, LOQ can reach 30fg/μL level. The preparation process of DNA Control is completely consistent with National Standard, therefore it has high purity and no protein and ion interference. DNA Control has been calibrated by National Standard 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 EC-D050T(50T)
B1	2XqPCR Mix	0.625mL
B2	Primer&Probe Mix	100μL
B3	DNA Dilution Buffer	2×1.5mL
B4	DNA Control (30ng/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℃ and it is recommended to be used within one year. B2 should be stored protected from light.
3	B2/B3/B4 can be stored at -20℃ for 2 years, while B1/B5/B6 can be stored at -20℃ for 1 year. B1/B5/B6 can also be purchased together as a separate set.

 

 

Quality Control on E.coli Host Cell DNA Residue Detection kit

Accuracy (Standard: ≦30%)	Intra Variation% 4.4-5.6
	Inter Variation% 3-15
Recovery%	53-81
Limit of Quantitation	30 fg/μL
Specificity	-

 

 

PCR Reaction System on E.coli 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.

 

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%.

 

No Template Control (NTC): In the reaction system, replacing the target template with DNA Dilution Buffer while keeping other components unchanged, and the Ct value obtained should be 'Undetermined' or Ct value≥35.