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Plasmid DNA Preparation .pdf



Original filename: Plasmid DNA Preparation.pdf
Title: Plasmid DNA Preparation
Author: http://www.synbio-tech.com/services/molecular-biology/plasmid-preparation/

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Plasmid DNA Preparation
For years, Synbio Technologies has provided comprehensive plasmid DNA
preparation services to our customers from research institutes and the
biotechnology and biopharmaceutical industries. We have developed and perfected
a manufacturing pipeline designed to meet the requirements of the Good
Manufacturing Protocol (GMP). These requirements guarantee both in-process
controls and final product quality control, to ensure that the highest quality product
will be generated. All the plasmids we manufactured are free of animal-derived
materials and can contain low levels of endotoxin (<0.005EU/μg, on request). This
process is designed to meet our customers’ diverse downstream applications such
as transfection, antibody preparation, and vaccine and gene-therapy research. In
addition to our services, we offer complete and detailed documentation that allows
us to track and monitor the whole manufacturing process from start to finish.
Throughout this process our project management team will monitor the whole
operation process to ensure delivery on time. This process culminates with extensive
quality control assays including sequence verification, restriction enzyme digestion
and endotoxin analysis. After passing our strict quality control assays, we guarantee
the quantity and quality of our delivered products. The result is high quality plasmid
DNA ready to ship to our customers all over the world. Additionally, to protect your
intellectual property, we are willing to sign an agreement or contract if deemed
necessary. We guarantee to protect our customers’ benefits and their products.
Dispensing is also available on your request; additional expense may be charged
depending on the quantity and other requirements.

Advantages
 Low Endotoxin Level:
Animal-derived materials free, <0.005EU/μg, on request

GMP-Like Manufacturing Pipeline:
Complete and detailed manufacturing documentation


Strict Quality Control:
ISO9001/2015 quality management




Fast procedure and on-time delivery

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Experimental Procedure

Service Specifications
Research
Grade

Application Grade

Endotoxin Level

NA

<0.005EU/μg

Quantity

0.1 mg to
gram level

1 mg to gram level

Turnaround time

Starting at 3
business days

Starting at 7 business days

Quality Control
Methods

Basic quality
control tests

Basic quality control tests plus Sterility assurance,
Restriction enzyme analysis and Sequence
verification

Standard Delivery
Package Contains

Prepared plasmid DNA, Certificate of analysis (COA) and QC
report

Note:Synbio Technologies provides services upstream of plasmid preparation such
as gene synthesis and subcloning into both commercial and custom vectors. This
allows us to offer a unique approach to satisfy all our customer’s requests.
Additionally, if you wish to provide the plasmid template, you will need to prepare
over 100 ng DNA sample (diluted with ddH2O or TE buffer), colonies (fresh) or
bacteria stored in glycerol.

Case Study
Application Grade Plasmid DNA Preparation
Method:
1. The target plasmid DNA was transformed into bacteria cells and a monoclonal
colony was selected to be cultured.
Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

2. Restriction enzymes were used to verify the sequence of the mini-prepped
plasmid.
3. Large volume bacteria culture or fermentation was performed to obtain a
sufficient amount of cell paste.
4. The cell paste was lysed to release plasmid DNA. The crude lysis solution was then
purified through several steps to dispose of impurities.
5. The bulk DNA solution was quality tested which included: restriction enzyme
digestion, sterility tests and endotoxin level tests.
6. After passing all of the quality control tests, the bulk DNA was formulated,
dispensed, labelled and finally delivered.
Quality Test Results:
1. Endotoxin Test: The negative control of endotoxin testing will not concrete, while
the positive control will concrete. These samples were not concrete (Fig.1), proving
that the endotoxin was successfully removed.

Fig. 1 Endotoxin testing, from left to right: negative control, positive control, sample,
endotoxin standard 5UR/mg
2. Sterility Test No bacterial colony was detected after 48 h culturing in
non-resistant medium (Fig.2).

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Fig. 2 Bacterial testing result comparison

Quality Control
Synbio Technologies plasmid preparation service focuses on quality. We have strict
quality control which can increase quality stability and reduce batch difference to
satisfy customers’ high-quality needs.
Quality Control

Standard Range

Measurement Technique

Appearance

Clear, transparent

Visual inspection

A260/280 Analysis

1.80~2.00

UV spectrophotometer

Concentration

Up to 5mg/ml, default 1±0.05
mg/ml

UV spectrophotometer

Supercoil Percentage
Analysis

>95%

Density scan on agarose gel

RNA Residue Analysis

Not detectable

Visual inspection on agarose gel

Genomic DNA Analysis

Not detectabl

Visual inspection on agarose gel

Sterility Assurance

No clone growth on LB
plate > 48h

Incubate 0.1mg of product on LB plate
for >48h

Restriction Enzyme Analysis As expected, no other minor Incubate with enzyme and analyze on
(optional)
band
agarose gel
Sequence Verification
(optional)

Correct

DNA Sequencing

Endotoxin Analysis
(optional)

<0.005EU/μg

Endotoxin Analysis Assay

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com

Related Services
 Syno®2.0 Gene Synthesis
 Syno®3.0 Gene Synthesis
 Synthetic DNA library
 PCR Cloning& Subcloning
 Site-Directed Mutagenesis
 Protein Expression and Purification

Contact us
Synbio Technologies
1 Deer Park Drive, Suite L-1
Monmouth Junction
NJ, 08852

Tel:+1 732-230-3003
Fax:+1 609 228 5911
Web:www.synbio-tech.com
E-mail:service@synbio-tech.com


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