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Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the analysis of RNase H cleavage products

✍ Scribed by Polo, Lenore M.; Limbach, Patrick A.

Publisher
John Wiley and Sons
Year
1998
Tongue
English
Weight
115 KB
Volume
33
Category
Article
ISSN
1076-5174

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✦ Synopsis

RNase H is an endonuclease which cleaves RNA at points of hybridization with DNA. However, certain ambiguities exist in terms of its speciÐcity and location of cleavage along the RNA strand. The analysis of RNase H reaction products of an oligoribonucleotide hairpin by matrix-assisted laser desorption/ionization time-of-Γ‘ight mass spectrometry (MALDI/TOF-MS) is demonstrated. The oligoribonucleotide studied has a highly stable secondary structure which reduces the efficiency of hybridization with the chimeric oligonucleotide used to direct RNase H cleavage. By monitoring the reaction products under di †erent conditions using MALDI/TOF-MS, the optimum variables for cleavage of this highly stable hairpin structure can be determined.

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