Related Experiment Video
Updated: Oct 20, 2026

Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
Published on: April 22, 2022
Identification of single stranded regions of DNA by enzymatic digestion with matrix-assisted laser
W P Bartolini1, C M Bentzley, M V Johnston
1Department of Chemistry and Biochemistry, University of Delaware, Newark 19716, USA.
Abstract:
Elucidating structure function relationships of DNA in cellular processes requires fast, reliable methods that can be applied to picomole amounts of sample. Higher order structure can be inferred by distinguishing paired and unpaired regions. It is shown here that enzymatic digestion coupled with product analysis by matrix-assisted laser desorption ionization (MALDI) is able to identify unpaired bases within structured DNA regions. The method is demonstrated with DNA duplexes having a five nucleotide mismatch as a 5' overhang, a 3' overhang, and an internal loop. Exo- and endonuclease digestions are performed under solution conditions (temperature, annealing, and enzyme buffers) which promote base pairing and specific enzyme activity. For each type of mismatch, the length and sequence of the single stranded region can be inferred from MALDI spectra taken as a function of digestion time.
More Related Videos
07:49Matrix-assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) Mass Spectrometric Analysis of Intact Proteins Larger than 100 kDa
Published on: September 9, 2013
09:20Identification of RNA Fragments Resulting from Enzymatic Degradation using MALDI-TOF Mass Spectrometry
Published on: April 11, 2022
Related Concept Videos
Matrix-Assisted Laser Desorption Ionization (MALDI)
MALDI-TOF Mass Spectrometry
Maxam-Gilbert Sequencing
Challenges of the Maxam-Gilbert Method
The...
Southern Blot
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
DNA Isolation
Sanger Sequencing