Navigation Links
Quantitation of DNA for Automated Sequencing Using the VersaFluor Fluorometer

standard curve was established for quantification of DNA concentrations ranging from 1005,000 g/ml by plotting seven points of varying DNA concentrations made by serial dilution of control calf thymus DNA. All dilutions were made in 2 ml of 1 g/ml Hoechst 33258 dye and included 0 ng DNA as a blank and 10,000 ng DNA as the upper range. The values were recorded in relative fluorescence units (RFUs) and were plotted on a graph with the x axis = DNA concentration (ng) and the y axis = RFU (Figure 2). The slope of the line was determined to be 1.0.

By employing the equation y = mx+b, we established the value of b to be equal to 0. Therefore, by setting the upper range value at 10,000 RFU using 10 l of 1 mg/ml calf thymus DNA and using 2 l of the sample DNA in 2 ml of the 1 g/ml Hoechst dye, the RFU reading is equal to ng/ml for our sample. We are then able to calculate the volume of DNA needed to obtain a 250 femtamolar concentration per reaction.


Results and Discussion
Using the Excel II sequencing kit (Epicenter) and quantifying our DNA with the VersaFluor fluorometer, we have been able to achieve highly readable and reproducible sequencing runs of up to 1,200 bp with the LiCor 4200 automated sequencer. This has enabled us to sequence the entire 2.2 Kb cDNA for Xenopus hypoxia inducible factor, xHIF, in several runs, and will allow us to sequence our genomic clone easily and efficiently.

The correct concentration of DNA in each reaction is the key to obtaining good reads on the LiCor 4200. This entails precise quantitation of the DNA. The VersaFluor fluorometer has proven to be easy to calibrate and provides excellent and reproducible results.

This work is
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Quantitation of Cabergoline at Extremely Low Plasma Concentrations with a Triple Quadrupole Mass Spectrometer
2. Bioanalytical Method Intraday Validation for the Quantitation of Paroxetine in Bovine Plasma using the TSQ Quantum Mass Spectrometer
3. Quantitation of the Protein MIA as a Marker for Chondrocytes in Research Samples
4. Comparison of Different Methods of mRNA Quantitation
5. Quantitation of RNA for Hybridization Blots Using the VersaFluor Fluorometer
6. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
7. Quantitation of GAD67 Gene Expression in Prefrontal Cortex of Schizophrenia Patients Using the iCycler iQ Detection System and Molecular Beacons, Rev A
8. Benchmark Plus Microplate Reader: Quantitation of Protein Concentration Using Two Different Colorimetric Assay Kits, Rev A
9. A Robust LC/MS/MS Method for the Quantitation of Very Long Chain Fatty Acids for Research into the Characterization of Peroxisomal Disorders
10. A new LC/MS/MS Research Method for Rapid Quantitation of Five Immunosuppressant Drugs, including Mycophenolic Acid (MPA)
11. Separation and Quantitation of R and S Enantiomers using Normal Phase Chromatography on an API 3000 LC/MS/MS System
Post Your Comments:
(Date:7/10/2014)... 2014 Product and process impurities ... abundance and are often “lost in the noise,” ... Join presenters Dr. Rowel Tobias, Senior Scientist, Protein ... Head of Quality at Nanotherapeutics, Inc., to lear ... speed detection and quantitation while achieving the required ...
(Date:7/10/2014)... (PRWEB) July 10, 2014 Terascala, the ... announced that Alan Swahn, a former vice president of ... team as vice president of marketing. Terascala’s software when ... and NetApp create the highest performance and most reliable ... appointed to guide Terascala’s channel expansion and broaden its ...
(Date:7/10/2014)... OMICS Group’s 5th International conference and exhibition ... at Double Tree by Hilton Beijing, China is all ... & Bioanalytical research methods only to facilitate improved living ... Dr. Srinubabu Gedela, MD of OMICS Group Inc has ... one in bringing a unique and international mix of ...
(Date:7/10/2014)... , July 10, 2014  Franciscan ... use of capnography for respiratory monitoring outside ... of healthcare leaders in embracing state-of-the-art patient ... effectively patients are breathing and can alert ... By measuring the amount of carbon dioxide ...
Breaking Biology Technology:Novel Ms Techniques Speed Detection of Biopharmaceutical Product and Process Impurities, New Webinar Hosted by Xtalks 2Terascala Expands Executive Team 2Terascala Expands Executive Team 3A Critical Review of Analytical Techniques for Improved Living Standards 2A Critical Review of Analytical Techniques for Improved Living Standards 3A Critical Review of Analytical Techniques for Improved Living Standards 4Franciscan St. Anthony Health-Crown Point Underscores Patient Safety Commitment Through Expanded Use of Capnography 2
... has succeeded in significantly increasing the sensitivity of the ... on this topic at Delft University of Technology on ... and astronomers from around the world are currently working ... of 66 advanced telescopes which will be placed at ...
... Iceland, February 3 deCODE genetics,(Nasdaq: DCGN ) ... letter from,the Nasdaq Listing Qualifications Panel (the "Panel") stating ... stock from the Nasdaq Global,Market to the Nasdaq Capital ... Wednesday, February 4, 2009. The Nasdaq Capital Market, which ...
... DHA (docosahexaenoic acid) omega-3 innovator and maker of the ... today that it has entered into a license agreement ... is a process widely used in food manufacturing that ... the nutritional value, shelf-life and flavor of the finished ...
Cached Biology Technology:Nanotechnology makes supertelescopes much more sensitive 2deCODE Announces Transfer of the Listing of its Common Stock from Nasdaq Global Market to Nasdaq Capital Market 2deCODE Announces Transfer of the Listing of its Common Stock from Nasdaq Global Market to Nasdaq Capital Market 3Martek, General Mills Collaborate on Microencapsulation Technology for Food Applications 2Martek, General Mills Collaborate on Microencapsulation Technology for Food Applications 3
(Date:7/11/2014)... There is evidence that under the normal ... and development, maintain neuronal environment, and exhibit ... and neurodegenerative disease. Previous studies have found ... cells after chemical induction have reduced viability, ... application. Prof. Xiaodong Yuan, Kailuan General Hospital, ...
(Date:7/11/2014)... that let nerve cells send out electrical signals, Johns ... tactic that may offer a new way to protect ... responsible way. , Their findingthat naturally occurring insect ... for a closely related onesuggests that insecticides can be ... like bees. A summary of the research, led by ...
(Date:7/10/2014)... 27, 2014  The American Academy of Forensic ... Standards and Technology (NIST) and the Department of ... and forensic science expert members to the Forensic ... element of the NIST,s Organization of Scientific Area ... first appointments made to the new FSSB that ...
Breaking Biology News(10 mins):Potent spider toxin 'electrocutes' German, not American, cockroaches 2Potent spider toxin 'electrocutes' German, not American, cockroaches 3Ten Members of the American Academy of Forensic Sciences Appointed to Forensic Science Standards Board (FSSB) 2Ten Members of the American Academy of Forensic Sciences Appointed to Forensic Science Standards Board (FSSB) 3
... we,ve been told, eliminating or severely limiting fats from ... in patients suffering from heart failure, a study at ... Results from biological model studies conducted by assistant professor ... researchers, demonstrate that a high-fat diet improved overall mechanical ...
... WASHINGTON The Chesapeake Bay Program is a ... jurisdictions in the bay watershed to oversee the restoration ... the extent of pollutants -- such as nitrogen, phosphorus, ... from the National Research Council assesses the framework used ...
... a researcher at the Royal Swedish Academy of Sciences, Margareta ... permafrost. "The changes we see are dramatic. And ... from the norm when compared with a longer term perspective", ... of the globe that is warming up fastest today. Measurements ...
Cached Biology News:Damaged hearts pump better when fueled with fats 2Chesapeake Bay program's 2-year milestones improve upon past strategies, but accounting of progress remains a challenge 2A new research report shows effects of climate change in the Arctic are more extensive than expected 2A new research report shows effects of climate change in the Arctic are more extensive than expected 3
... has over 23 years of experience in ... have made antibodies to large and small ... antigens, and plasmid DNA-encoded antigens. We understand ... so our hybridoma development programs are never ...
... Rapid-Screen cDNA Library Panels were designed for ... sets of PCRs to identify the desired ... performed in a 96-well "Master Plate," where ... clones. Having identified the positive well(s) by ...
... The Rapid-Screen cDNA Library Panels were ... just three sets of PCRs to identify ... analysis is performed in a 96-well "Master ... from 5,000 clones. Having identified the positive ...
... CBED Maxi Kit provides a fast and ... protein solutions for buffer exchange, and for removing ... kit is based on spin-column chromatography using Norgens ... such as MgCl 2 , NaCl, KCl, CaCl ...
Biology Products: