Sequence Analysis Using Base-Specific Cleavage and MALDI-TOF MS

COMPARATIVE SEQUENCE ANALYSIS USING BASE-SPECIFIC CLEAVAGE AND MALDI-TOF MS 

The completion of the Human Genome Project, however, does not imply that there remains no further need for sequencing. Quite to the contrary, the availability of a genetic blueprint allows for large-scale genetic variability studies. This requires efficient sequencing technologies. 

The first step of this process involves PCR amplification of the target region. The use of a primer that carries a T7 promoter sequence at its 5 end enables incorporation of a transcription start site in the PCR product. This allows for in vitro transcription of the PCR product into a single-stranded RNA molecule in a subsequent step. Endonucleolytic cleavage of the RNA molecule is then performed by adding a ribonuclease. RNase A cleaves pyrimidine residues (Cytosine and Uracil) and therefore it is not 

MALDI-TOF MS acquisition is a unique combination that will play an increasingly important role in the discovery of novel sequence variations. 

The analysis of epigenetic changes, mainly the covalent addition of methyl groups to the 5 position of Cytosine in CpG sequences, has become an increasingly important field in genetic research. The DNA methylation has 

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