Large Scale Plasmid DNA Purification and Production (100mg-multi gram)
Low Endotoxin Plasmid DNA Purification and Processing

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

NTC is an industry-leading provider of highly-purified plasmids, having highest yielding fermentation process

(greater than 2.6g/L) and purification. The process is scalable, and is being used in GMP clinical grade DNA production.

Large scale DNA plasmids at NTC are grown in a fed-batch fermentation process, permitting control over feed rate, pH, temperature, and dissolved oxygen. NTC’s patented HyperGRO™ fermentation process and media has produced record pDNA yields of >2.6g/L, and typically produce 0.5-1.5g/L using a high copy pUC, or similar origin, and a recA-, relA-, endA- host strain, such as DH5 alpha. Highly concentrated HyperGRO™ plasmid preparations naturally result in cleaner, more highly-purified DNA (due to dramatically reduced biomass), and more cost-effective production as well. (See related molecular services below)


At no additional charge,
Each endotoxin-free DNA prep includes:

1) Transformation
2) Glycerol stock archiving
3) Lot sample archive
4) Custom formulation
5) Certificate of Analysis (C of A)

Additional Testing
Advantages of NTC’s Plasmid Manufacturing Capabilities

• Highest yields (>2.6g/L)

• Highest quality purification (>95% supercoiled plasmid DNA)

• Lowest biomass process (5% pDNA, dry mass)

• Lowest cost per unit DNA

• Lowest chemical use for "greenest" manufacturing

• Choice of antibiotic-free or Kanamycin R

• Vector to process matching for highest overall performance

• Reduced metabolic burden for production of toxic or unstable plasmids

• Tech transfer for process continuity (regulatory)
Production of highly-purified research-grade plasmid DNA at NTC’s Lincoln, Nebraska, USA plant range from 1mg to multi-gram scale, and production capabilities for GMP-grade DNA at NTC’s CMO partners range from a hundred milligrams upward.

XXEasy to Order:

NTC is an industry leading provider of highly-purified plasmids. Ordering plasmids from NTC is a fast and easy 2 step process, and there is no where else you will receive the attention to detail and great service for the cost.

Timeline: First time orders require 2-3 weeks. Reorders can be from 1 day to two weeks, depending upon the arrangements made (just in time re-ordering is by prior agreement).

1) Call NTC for a quotation, (888) WIZ-BANG. Best pricing is available based on quantity ordered and copy number. Most high-copy plasmids have a pUC origin of replication, or equivalent, and result in the lowest cost. NTC is a longtime industry leader in quality and cost-effectiveness.


2) Send NTC 1microgram of your plasmid DNA, spotted on Whatman 3MM or FTE filter paper, together with NTC's Plasmid DNA Order Form filled out. (The form is designed to prompt you for all information needed for propagation and testing.)

Plasmid DNA Order Form

XXNTC will:


• Transform the production host.

• Prepare a glycerol stock using "low metabolic burden technology"

• Grow, and purify the desired quantity of pDNA, using endotoxin-free reagents and procedures. 

• At no additional cost, NTC will also formulate the DNA according to specification (default is 0.5mg/ml in TE buffer). 

• A large-scale certificate of analysis will be provided, including: gel photo (digested and undigested DNA, ladder); concentration and quantity; information and testing for residual: RNA, genomic DNA (qPCR). Upon request, additional testing for protein, endotoxin [LAL assay] and bioburden are available .(see table below) 

XXXXX·  Spectrophotometric analysis (A260/280)
XXXXX·  Quantity
XXXXX·  Concentration (usually 0.5mg/ml, in TE buffer)
XXXXX·  Volume
XXXXX·  Gel photo
XXXXX·  Residual RNA (gel assay)
XXXXX·  Residual genomic E. coli DNA (qPCR)
XXXXX

• A large-scale certificate of analysis will be provided including gel photo (digested and undigested DNA, ladder), concentration and quantity

• In addition, NTC will retain an archival lot sample, and a glycerol stock of your plasmid for future reorders (at least one year retention). 
XXNTC's testing at no additional charge and cost comparison with NTC's competitors





XXAdditional testing is available upon request

Additional testing

Residual Protein test (modified BCA)--small-scale pDNA prep
NTC-RPSS-RPA
$75.00

Residual RNA test (spiked gel)--small-scale pDNA prep
NTC-RPSS-RRNA
$75.00

Genomic DNA test (RT-PCR)--small-scale pDNA prep
NTC-RPSS-GDNA
$150.00

Endotoxin test--small-scale pDNA prep
NTC-RPSS-ENDO
$50.00
0.2 um filtration of purified plasmid
NTC-RPSS-FILTR
$50.00

Bioburden test--small-scale pDNA prep
NTC-RPSS-BBDN
$50.00
Residual Protein test (modified BCA)--large-scale pDNA prep NTC-RPLS-RPA
$75.00
Residual RNA test (spiked gel)--large-scale pDNA prep NTC-RPLS-RRNA
$0.00
Genomic DNA test (RT-PCR)--large-scale pDNA prep NTC-RPLS-GDNA
$0.00
Endotoxin test--large-scale pDNA prep NTC-RPLS-ENDO
$0.00
Bioburden test--large-scale pDNA prep NTC-RPLS-BBDN
$50.00

XXRelated molecular services:

1) Precision Cloning

2) Strain development and engineering by NTC’s Genome Mass Transfer (GMT™) technology, for precise knock-ins and knock-out assemblies in E. coli host strains

3) Process Development / Engineering

4) Recombinant Proteins


Other DNA production services available at NTC:

·       E. coli cell paste containing HyperGRO™ enriched plasmid

·       HyperGRO™ media kits for fed-batch plasmid DNA fermentation

·       Site-license for in-house use of NTC’s HyperGRO™ process

·       Small scale plasmid DNA preparations;

·       Linear vectors (digested, gel-purified, highly-competent for cloning).