Navigation Links
Automation of TempliPhi and GenomiPhi DNA Amplification Kits on the Genesis Freedom Robotic Sample Processor

M. Adams and R. Deadman
GE Healthcare, Piscataway, NJ, US


GE Healthcare offers a range of DNA amplification products to meet all genomic DNA and sequencing template preparation needs. A combination of these kits with the Genesis Freedom™ Robotic Sample Processor (RSP) from Tecan (Fig 1) creates a total solution for high-throughput DNA preparation.

Traditional DNA purification methods often use organic solvents, are difficult to automate, or produce variable yields. The minimal number of liquid transfers in the TempliPhi™ and GenomiPhi™ DNA amplification procedures are easy to automate, with each reaction producing the same amount of high quality DNA. TempliPhi DNA Amplification Kit and TempliPhi Large Construct DNA Amplification Kit are used to prepare small (plasmids/M13) or large (fosmids/BACs) circular DNA templates, respectively, for sequencing. GenomiPhi DNA Amplification Kit prepares genomic DNA from various sources that can be directly used for genetic analyses. The flexibility of the Genesis Freedom workstation also allows easy reaction setup for downstream applications such as sequencing or PCR.


DNA amplification
TempliPhi DNA Sequencing Template Amplification Kit offers the simplest solution to plasmid DNA preparation. Microgram quantities of DNA can be prepared from picogram amounts of purified DNA, colonies, or bacterial cultures using Phi29 DNA polymerase and rolling circle amplification. This unique approach removes the need for culture growth, resulting in high quality DNA templates with less than 20 min of hands-on time.

Amplified DNA can be used directly for sequencing with any chemistry and sequencing platform. TempliPhi Large Construct Kit has been specifically optimized for amplifying large constructs such as fosmids or BACs for sequencing. Uniform quantities of DNA are amplified in all samples, eliminating the need for quantitation.

GenomiPhi DNA Amplification Kit representatively amplifies single- or double-stranded linear genomic DNA using Phi29 DNA polymerase to perform strand displacement amplification. Microgram quantities of representative DNA are produced from small amounts of starting material that can be used immediately in downstream applications.

For all kits, DNA amplification is achieved by combining the starting DNA with random hexamer primers, Phi29 DNA polymerase, dNTPs, and buffer components optimized for DNA synthesis. This reaction mixture is incubated overnight at 30 °C, during which time the available nucleotides are consumed and converted into high molecular weight fragment copies of the input DNA.


Automation
The amplification protocols are simple and allow straightforward automation. The Genesis Freedom series of RSPs from Tecan are ideal for automating DNA amplification reactions and downstream applications. The arm of the Tecan liquid handler is equipped with four or eight tips and can perform all required liquid handling steps with high sensitivity and accuracy. When the amplification reaction stops because the nucleotides in the reaction are exhausted, DNA yields are equivalent across all samples (Fig 2). Disposable tips eliminate cross-contamination concerns. With an eight-channel liquid handling arm, multiple 96-well plates of TempliPhi or GenomiPhi DNA amplification reactions can be prepared in half the time of manual pipetting. Whatever the post-amplification application (Fig 3), the open platform can be adapted. The Tecan Gemini software system allows easy protocol setup and operation of the workstation.



Conclusions
The combination of GE Healthcare’s DNA amplification products and Tecan’s Genesis Freedom workstation produces a high-performance platform for automating the labor-intensive steps traditionally associated with manual methods of DNA preparation. The simplicity of the protocols and the robotics system allow a short setup time. Amplification reactions produce consistent, high quality DNA across all samples, eliminating time-consuming quantitation and purification steps. DNA can be used directly in downstream applications with no further purification.

For more information, including protocols, please refer to the application note Automation of TempliPhi and GenomiPhi DNA Amplification Kits on the Genesis Freedom Robotic Sample Processor (63-0056-16) which is available at: www.amershambiosciences.com/promo_dm203



back to top
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. RNAqueous-96 RNA Isolation Kit Automation Protocol
2. Scalable Automation of the TempliPhi DNA Sequencing Template Amplification and DYEnamic ET Terminator Sequencing Kits on the Sciclone ALH 3000
3. Automation of the Eppendorf Perfectprep BAC 96 DNA Kit on the Caliper Life Sciences Sciclone ALH 3000
4. Oragene and Whole Genome Amplification with GenomiPhi
5. GenomiPhi: protocol for BAC amplification and fabrication of slides for array CGH
6. GenomiPhi amplification of FFPE tissue DNA for array CGH
7. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
8. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
9. Greater Amplification Specificity with New Hot Start PCR Enzyme
10. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
11. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
Post Your Comments:
(Date:9/19/2014)... -- Pfenex Inc. (NYSE MKT: PFNX), a clinical-stage biotechnology ... to manufacture proteins including biosimilar therapeutics, today announced that ... Annual NewsMakers in the Biotech Industry conference in ... chief executive officer of Pfenex, will provide an overview ... Friday, September 26, 2014 at 10:00 a.m. EDT/7:00 a.m. ...
(Date:9/19/2014)... September 19, 2014 ... http://www.researchandmarkets.com/research/b2jxhs/micro_market ) has announced the addition of the  ...  report to their offering.       (Logo: ... one of the fastest-growing segments in the chromatography ... in 2013, and expected to reach $2.0 billion ...
(Date:9/18/2014)... HealthTronics, Inc ., a leading ... and services, announced that a poster related to ... external beam radiotherapy (EBRT) is being presented at ... (MAAUA) Annual Meeting in Baltimore, Maryland September 18-21, ... and External Beam Radiation as Treatments for Localized ...
(Date:9/18/2014)... Mich. , Sept. 18, 2014  Health Enhancement Products, ... firm investigating early-stage algae bioactive compounds and metabolic processes, is ... the company,s board of directors. Ms. Nola E. Masterson ... investment advisor joined the board effective September 17, 2014. ... Masterson agree to join our board," states Andrew Dahl ...
Breaking Biology Technology:Pfenex to Present at Upcoming Industry Conference on September 26 2Micro Market Monitor : Global Pre-packed Chromatography Columns 2HealthTronics Announces Data Comparing Cryotherapy Versus Radiation for Treating Prostate Cancer Poster Presented at the American Urological Association Regional Meeting 2Health Enhancement Announces New Board Member In Advance Of Annual Shareholder Meeting 2
... Inc. (Nasdaq: ALTH ), a biopharmaceutical company ... therapeutics for,the treatment of cancer, today announced that ... corporate overview at the Bear Stearns 20th Annual,Healthcare ... will take place at 9:00 a.m. (Eastern) on,Monday, ...
... 4 Medarex, Inc.,(Nasdaq: MEDX ) announced today ... 20th Annual Healthcare Conference at 9:00 AM Eastern Time ... at The Grand Hyatt,New York in New York City. ... for replay through the Webcast page in the Investor,Relations ...
... Statistical Considerations for Model, Selection and Bias; Dr. Klaas Prins ... ... Using Metadata, MOUNTAIN VIEW, Calif., ... provider of software and,strategic services for optimizing clinical drug development, ...
Cached Biology Technology:Allos Therapeutics to Present at the Bear Stearns 20th Annual Healthcare Conference 2Medarex to Present at the Bear Stearns 20th Annual Healthcare Conference 2Pharsight to Present at ROeS Seminar in Bern, Switzerland on September 10 & 11, 2007 2Pharsight to Present at ROeS Seminar in Bern, Switzerland on September 10 & 11, 2007 3
(Date:9/18/2014)... Boulder, Colo., USA Miranda, a small, icy moon ... and enigmatic bodies in the solar system. Despite its ... episode of intense resurfacing that resulted in the formation ... -- polygonal-shaped regions called coronae. , These coronae ... is at least 200 km across. Arden corona, the ...
(Date:9/18/2014)... "frequent flyers" program, fruit flies surely would be eligible ... increasing numbers of fruit fly research studies. One fruit ... and another is scheduled launch to the station Sept. ... December. , Fruit flies are biomedical research models studied ... organisms can reveal the basis for health and disease ...
(Date:9/18/2014)... stored in DNA, tiny strands of nucleic acid that ... express this genetic data, our DNA is copied into ... that perform tasks in our cells. , Several ... molecule. Unlike all other known RNAs, this molecule is ... are abundant, little has been known about how they ...
Breaking Biology News(10 mins):Miranda: An icy moon deformed by tidal heating 2Miranda: An icy moon deformed by tidal heating 3Miranda: An icy moon deformed by tidal heating 4Miranda: An icy moon deformed by tidal heating 5Miranda: An icy moon deformed by tidal heating 6Miranda: An icy moon deformed by tidal heating 7Buzzing with activity: Fruit flies orbit Earth for science 2Buzzing with activity: Fruit flies orbit Earth for science 3Buzzing with activity: Fruit flies orbit Earth for science 4Scientists find how mysterious 'circular RNA' is formed, claim muscular dystrophy link 2
... A study of DNA rearrangements in roundworm chromosomes may offer ... tumors. A report of the research led by University ... was published in the April 22 online edition of the ... region of repetitive DNA sequence that protects the ends of ...
... how a single-celled fungus survives in low-oxygen settings may ... organ transplants or AIDS. A report on the ... 22 in Molecular Cell . Previous work ... model organism that,s often used to study individual genes, ...
... in German . It would appear reasonable ... similar genetic blueprints. However, scientists from the Max Planck Institute ... international research team have now decoded, for the first time, ... lyrata ), a close relative of the thale cress ( ...
Cached Biology News:Study in roundworm chromosomes may offer new clues to tumor genome development 2Simple fungus reveals clue to immune system protection 2Evolution can cause a rapid reduction in genome size 2Evolution can cause a rapid reduction in genome size 3
Request Info...
Tight junctions are complexes of proteins that create intercellular boundaries between the plasma membrane domains of epithelial and endothelial cells. Antigen: Synthetic peptide corresponding to...
TNF-Related Apoptosis Inducing Ligand-Receptor-1...
NATIVE PORCINE COLIPASE...
Biology Products: