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NTC Contacts:
Phone: (402) 323-6289 (natx)
Fax: (402) 323-6292
Email: support@natx.com

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NTC PRODUCTS AND SERVICES

MAMMALIAN EXPRESSION VECTORS
X• Antibiotic-Free Vectors
X• Rig-I Activating Vectors
X• Linear Vectors
X• Recombineering Vectors
X• Gene Silencing (transient)
X• Gene Silencing (retroviral, stable)
X• Anti-Silencing Elements
X• Vector Pricing

CELL LINES
X• Autolytic E. coli Cell Lines
X• dcm- Strains

VACCINES & ADJUVANTS
X• DNA Vaccines
X• Rig-I Activating DNA Vaccines
X• Thermostable Protein Expression

PRECISION CLONING
X• Services and Pricing

RECOMBINANT PROTEINS
X• pVEX™-E. coli Expression Vectors
X• Fluorescent Proteins
X• Products, Vectors, Services, Prices
X• CofA (SAMPLE)

PLASMIDS
X• Small Scale Plasmid DNA
X• Small Scale C of A
X• Large Scale Plasmid DNA
X• Large Scale C of A
X• Plasmid DNA Order Form
X• E. coli Cell paste
X• NEW! Plasmid Detox Service
X• Additional Testing

PROCESS DEVELOPMENT

DNA FERMENTATION PROCESS
X• HyperGRO™ Publications
X• E. coli Cell Paste

HYPERTAQ™ DNA POLYMERASE

CONTRACT QC
X• E. coli Genomic DNA Testing
X• Residual RNA qPCR
X• Residual RNA gel
X• Supercoil Assay
X• Lymulus Endotoxin Testing

Contract QC (Single Samples)
Plasmid DNA Homogeneity/Supercoil Assay by Agarose Gel Electrophoresis
 NTC-CQC-SUPAGE1
$75.00
Quantitation of Residual DH5alpha E.coli RNA by RT-qPCR Using Taqman Technology (GLP) NTC-CQC-RNA1
Custom Assay
Inqure
Quantitation of Residual DH5alpha E.coli DNA Using Taqman Technology (GLP) (1 sample) NTC-CQC-DNA1
$150.00
Endotoxin testing (1 sample) NTC-CQC-ENDO1
$80.00
Residual RNA test by Agarose Gel Electrophoresis (1 sample) NTC-CQC-RNAGEL1
$75.00


XXXE. coli Genomic DNA Testing

Step One qPCR machine, from Applied Biosystems DNA plasmids are grown in E. coli host cells, and subsequent purification generally results in some degree of contamination of plasmid DNA preparations with bacterial DNA. Quantitative PCR (qPCR) has become the generally accepted method for determination of residual genomic DNA in plasmid DNA.

Fortunately, repetitive sequences in the E. coli genome make it relatively easy to quantitate the levels of genomic DNA, over at least five logs of concentration (fig. 1, A&B).  Testing involves comparison of a standard curve with samples.
qPCR testing for genomic DNA is an option with each DNA preparation of 100mg or greater.  However, it can also be added onto the cost of a small scale preparation ($150).
Amplification plot, showing the relative abundance of DNA standards in qPCR
Fig. 1A Standard curve amplification plot:
genomic DNA
Standard curve, showing excellent linearity of DNA standards used in qPCRFig. 1B. Standard curve (red) with samples (blue):
genomic DNA
For guidance, the FDA has recommended a minimum level of 1% genomic DNA for DNA vaccines that are used in humans[1].
[1] Guidance for Industry: Considerations for plasmid DNA vaccines for infectious disease indications, 2007.




XXLymulus Endotoxin

Endotoxin (LPS) Testing - LAL Kinetic Assay - Lymulus (horseshoe crab) amoebocyte lysate (LAL) test

The Lymulus amebocyte lysate (LAL) kinetic assay is a definitive test that is used for the detection and quantitaton of endotoxins (aka: lipopolysaccharides [LPS], or endogenous pyrogens) produced by gram negative bacteria, that may be present in biotechnological products (especially fermentation products). If your biological materials are being used in animal studies, or in highly sensitive cell culture systems, you will want to know the amount of endotoxin present, which could dramatically affect your results. Prime targets for endotoxin testing include recombinant plasmids and proteins (especially those being grown in E. coli or other gram negative organisms), water, buffers, serum components and other materials that are being tested in vitro or in animals. NTC's kinetic assay compares your product against current international standards, and provides a statistical analysis of the product, including 'spiked' samples to determine possible masking of endotoxin detection.