Molecular Biology
Preparation of Digoxigenin labeled riboprobes
Preparation of riboprobes useful for in situ hybridization
Inverse-PCR (I-PCR) for the identification of the breakpoints of deletions
The Inverse-PCR is a method that allows to amplify unknown regions of DNA, starting from the flanking known region on which are positioned the primers, in inverse direction compared to a normal PCR. It is necessary circularizing the DNA fragments, using ligase, then perform the I-PCR.
Circularization of DNA fragments for the molecular characterization of new deletions
During the molecular characterization of new deletions (to define the breakpoints) can occur that one extreme is known, but not the second one. In these cases it is very useful and suitable to perform the circulatization by ligase of DNA fragments containing the beakpoints of the deletion, followed by the inverse-PCR. Using these methods we were able to define the breakpoints of two new beta-Thalassemia deletions.
