Interfacing an Electrodynamic Ion Funnel with an Electrospray Ionization, Orthogonal Extraction, Time of Flight Mass Spectrometer for Improved Transmission of Single Stranded DNA Molecules.
EMSL Project ID
1931
Abstract
In sequencing the Human Genome, the current technology of choice for separating Sanger DNA sequencing reactions is gel electrophoresis. This technique provides high resolution with single base separations possible out to 1000 base pairs. However, the time required to separate reactions by this method is on the order of several hours. One possible means of reducing the separation time is to replace the gel electrophoresis with time of flight mass spectrometry (TOF-MS). This method easily provides the mass resolution required to separate the individual fragments and determining the DNA sequence. The time required to resolve the fragments by TOF-MS is on the order of several hundred microseconds. Early work by Lloyd Smith, University of Wisconsin-Madison, and others have shown that the current utility of TOF-MS for sequencing is limited by two factors, over all sensitivity and reduced sensitivity with increasing mass/fragment size. We plan to investigate the utility of the Electrodynamic Ion Funnel developed under Richard Smith, Pacific Northwest National Laboratory, to improve the ion transmission efficiency/sensitivity of an electrospray ionization TOF-MS. To complete this work we will need to duplicate the Ion Funnel and interface it with our TOF-MS instrument in Madison Wisconsin. We will be collaborating with the Harold Udseth, Pacific Northwest National Laboratory, on the construction of the funnel. This will involve a visit or series of visits to EMSL to discuss construction requirements. Once we have constructed the apparatus, Harold Udseth will guide us through the implementation of the funnel via telephone or email. The effort will not require any of the instrumentation facilities of ESML.
Project Details
Project type
Exploratory Research
Start Date
2000-03-15
End Date
2000-03-16
Status
Closed
Released Data Link
Team
Principal Investigator