Navigation Links
GenomiPhi amplification of FFPE tissue DNA for array CGH

Cy3- and Cy5-labeled dCTP and Klenow fragment. Cy3- and Cy5 labeled probes were separated on a 2% agarose gel for size analysis and imaged using a Typhoon 8600 Variable Mode Imager. Hybridization was performed on SpectralChip 2600 BAC Arrays according to the manufacturer’s recommended conditions. The arrays were scanned on a GenePix 4000B scanner and signal intensities quantitated using GenePix Pro 3.1 software.


Results
DNA extraction from FFPE tissue
DNA was extracted from human liver FFPE tissue slides using a Chelex-100 resin extraction method. The extraction resulted in genomic DNA with an average yield of 800 ng from 20-µm specimen slides. The quality of DNA was verified using conventional PCR, demonstrating amplification of specific human gene fragments from 150 to 1100 base pairs in length (5).

Amplification of FFPE DNA
DNA extracted from FFPE tissue slides was amplified using GenomiPhi DNA Amplification Kit and DOP-PCR. Approximately 10–25 ng of FFPE-extracted DNA was used for each amplification.

When DOP-PCR amplification was carried out on FFPE- isolated DNA using three primers (DOP1, DOP2, and DOP3) as described in the Sanger protocol, only the DOP2 primer was successful in amplifying FFPE template DNA (Fig 1). The sizes of amplified DNA from DOP-PCR amplifications were small, with fragments ranging from 100 to 600 bases. The amplification was found to be dependent on the quality of FFPE DNA, resulting in lower yields than the GenomiPhi method (Table 1).


In contrast, GenomiPhi DNA Amplification Kit proved more consistent and reliable in amplifying FFPE-extracted DNA (Fig 2). The success of GenomiPhi amplifications was measured by the presence of a smear on the agarose gel indicat
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10

Related biology technology :

1. Oragene and Whole Genome Amplification with GenomiPhi
2. GenomiPhi: protocol for BAC amplification and fabrication of slides for array CGH
3. Automation of TempliPhi and GenomiPhi DNA Amplification Kits on the Genesis Freedom Robotic Sample Processor
4. The TripleMaster PCR System for three different amplifications
5. Successful PCR amplification and subcloning of a GC-rich DNA fragment
6. Advantages of Roche Applied Science amplification products
7. Quality assurance of RNA derived from laser microdissected tissue samples obtained by the PALM MicroBeam System using the RNA 6000 Pico LabChip kit.
8. Protocol for GenomePlex Whole Genome Amplification from Formalin-Fixed Parrafin-Embedded (FFPE) tissue
9. 2D-LC analysis of phosphopeptides in brain tissue using Ettan MDLC and Finnigan LTQ
10. Sensitive identification of phosphopeptides in brain tissue using Ettan MDLC and Finnigan LTQ
11. High sensitivity quantitation of metabolites of nitrofuran antibiotics in animal tissue using LC/MS/MS
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: GenomiPhi amplification FFPE tissue DNA for array CGH

(Date:12/17/2014)...  United Therapeutics Corporation (NASDAQ: UTHR ) ... changes to Martine Rothblatt , Ph.D,s compensation program. ... United Therapeutics announced the promotion of Roger Jeffs ... David Zaccardelli , Pharm.D. to Executive Vice President ... Dr. Rothblatt,s title will change from Chairman and Chief ...
(Date:12/15/2014)... NEW YORK & PETACH TIKVAH, ... Therapeutics Inc. (NASDAQ: BCLI ), a leading ... announced today that wholly-owned subsidiary Brainstorm Cell Therapeutics Ltd. ... Israel,s Office of the Chief ... has been in receipt of grant support from the ...
(Date:12/13/2014)... CanDiag, Inc., a University of North Carolina at ... to OncoTab, Inc. , “The new name is ... said CEO and Chief Science Officer, Dr. Pinku Mukherjee, ... diagnostics to include imaging and therapies, and our new ... in 2011 by Dr. Mukherjee, an Irwin Belk Endowed ...
(Date:12/13/2014)... (PRWEB) December 13, 2014 The ... the major market drivers, threats, opportunities, and challenges. ... by three major players, namely, Thermo Fisher Scientific, ... (Netherlands). These players jointly accounted for approximately ~65% ... , Full Copy for This Report @ ...
Breaking Biology Technology:United Therapeutics Corporation Announces Executive Promotions and CEO Compensation Change 2United Therapeutics Corporation Announces Executive Promotions and CEO Compensation Change 3United Therapeutics Corporation Announces Executive Promotions and CEO Compensation Change 4BrainStorm Receives $1,100,000 for its 8th Annual Non-Dilutive Grant from Israel's Chief Scientist Office 2BrainStorm Receives $1,100,000 for its 8th Annual Non-Dilutive Grant from Israel's Chief Scientist Office 3CanDiag Inc. Expands Product Scope Under a New Name: OncoTab, Inc. 2Synthetic Biology Market to HIt $5.6304 Billion by 2018 - New Research Report by MarketsandMakets 2Synthetic Biology Market to HIt $5.6304 Billion by 2018 - New Research Report by MarketsandMakets 3
... Dynatronics,Corporation (Nasdaq: DYNT ) will release ... June 30, 2007 during the morning of Friday, ... call for investors later that,day at 1:00 p.m. ... and fiscal year.,Those interested in participating should call ...
... Leadership in Clean Technology and Global Business Potential, ... Solazyme, Inc., a,leading algal biotechnology company, today announced ... of the GoingGreen 100 Top Private Companies for,2007. ... by the,AlwaysOn editorial panel. In order to be ...
... ACCESS PHARMACEUTICALS, INC.,(OTC Bulletin Board: ACCP) announced ... BusinessWeek, highlighted Access Pharmaceuticals in the,most recent ... is also available,on http://www.BusinessWeek.com under ... article focused upon the major opportunity Access, ...
Cached Biology Technology:Solazyme, Inc. Selected by AlwaysOn as a GoingGreen 100 Top Private Company Award Winner 2Solazyme, Inc. Selected by AlwaysOn as a GoingGreen 100 Top Private Company Award Winner 3Access Highlighted in Recent BusinessWeek Article 2
(Date:12/17/2014)... N.C. , Dec. 16, 2014 Valencell, ... licensed its PerformTek biometric technology to industry leaders such ... highly accurate, clinically validated, biometric wearable products. These products ... 6-9 in Las Vegas . ... technology is accurate, flexible and robust – with the ...
(Date:12/11/2014)... , Dec. 09, 2014 Research and Markets ... ( http://www.researchandmarkets.com/research/9ql3kr/biometrics_market ) has announced ... India 2015-2019" report to their offering. ... market is the adoption of multimodal biometric systems. ... of an individual for verification and identification purposes. ...
(Date:12/10/2014)... Dec. 9, 2014  Wake Forest Baptist Medical Center today ... its School of Medicine. Funding for this $50 million ... that will be publicly launched next summer. ... the former 60 series R.J. Reynolds Tobacco Company complex, ... will begin immediately with plans to be ready to ...
Breaking Biology News(10 mins):Valencell PerformTek Biometrics Power the Most Accurate Wearables at CES 2015 2Biometrics Market in India 2015-2019: Key Vendors are 3M Cogent, NEC, Safran and Suprema 2Wake Forest Baptist to Build New Medical Education Facility In Wake Forest Innovation Quarter 2Wake Forest Baptist to Build New Medical Education Facility In Wake Forest Innovation Quarter 3Wake Forest Baptist to Build New Medical Education Facility In Wake Forest Innovation Quarter 4
... The first-ever drug known to prevent some preterm births, which won ... by the March of Dimes. "For the first time, we ... baby too soon, giving them hope that their next child will ... Alan Fleischman, MD, senior vice president and medical director of the ...
... not always what they seem when it comes to fishsomething ... are finding out. Using modern genetic analysis, combined with traditional ... thought to be three species of blenny in the genus ... findings are published in the scientific journal ZooKeys , ...
... hunt by Johns Hopkins scientists for actively "jumping genes" in ... but static, contains numerous pesky mobile elements that may help ... traits and disease risks. Using bioinformatics to ... the reference human genome to raw whole-genome data from 310 ...
Cached Biology News:March of Dimes welcomes FDA approval of progesterone injections 2Smithsonian scientists discover 7 new species of fish 2Johns Hopkins researchers capture jumping genes 2
... The Zero Blunt ... offers the fastest and ... (greater than or equal ... PCR products amplified with ...
Blot Paper 7 x 8.5cm semi-dry western blots (order number RB-400)...
... Create 100% capped transcripts with the ... higher yields of 100% capped (versus 40% ... mRNA messages in about 2 hours. mScript™ ... has been shown to be up to ...
Immunogen: Synthetic peptide: M(1) C V P R S G L I L S(11) Storage: -20 C, Avoid Freeze/Thaw Cycles...
Biology Products: