Use of paraffin wax film in MALDI-TOF analysis of DNA.
Poor detection limits and strong salt effects are two of the main problems encountered in the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometric analysis of DNA. This work demonstrates that a probe tip with a paraffin wax film (Parafilm) surface improves the MALDI performance in DNA analysis compared to the commonly used metal surface. First, the use of Parafilm increases the detection sensitivity. It was found that the detection limit achieved with Parafilm was 5 times better than that obtained using stainless steel for a 85mer. More importantly, the Parafilm method could improve detection of larger DNA components in the presence of a large excess of a smaller DNA component or in a DNA mixture. This feature is important to analyses of PCR and sequencing products. Second, we found that the use of Parafilm increased the salt tolerance limits for the 17-, 41-, and 85mers studied in this work and that the salt effect was less sensitive to the DNA size. Third, this method offers other analytical benefits, including producing a more homogeneous coverage of matrix/DNA, adding no extra cost and time to sample preparation, and eliminating the commonly required step for cleaning the probe after analysis. In this paper, we will also present our perspectives on why the use of Parafilm can improve the MALDI-TOF performance in DNA analysis.
Department of Chemistry, Cleveland State University, Ohio 44115, USA.
K C
KC
Hung
H
H
Rashidzadeh
Y
Y
Wang
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B
Guo
D004247:DNA / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D016612:Paraffin Embedding / Q000379:methods*
D004247:DNA