10.6084/m9.figshare.5122642.v1 Morozkin E.S. Morozkin E.S. Loseva E.M. Loseva E.M. Karamysheva T.V. Karamysheva T.V. Babenko V.N. Babenko V.N. Laktionov P.P. Laktionov P.P. Vlassov V.V. Vlassov V.V. Rubtsov N.B. Rubtsov N.B. Supplementary Material for: A Method for Generating Selective DNA Probes for the Analysis of C-Negative Regions in Human Chromosomes Karger Publishers 2011 C-negative regions Linker-adapter polymerase chain reaction (LA-PCR) FISH Supernumerary marker chromosomes 2011-07-28 00:00:00 Dataset https://karger.figshare.com/articles/dataset/Supplementary_Material_for_A_Method_for_Generating_Selective_DNA_Probes_for_the_Analysis_of_C-Negative_Regions_in_Human_Chromosomes/5122642 Linker-adapter polymerase chain reaction (LA-PCR) is among the most efficient techniques for whole genome DNA amplification. The key stage in LA-PCR is the hydrolysis of a DNA sample with restriction endonucleases, and the choice of a restriction endonuclease (or several endonucleases) determines the composition of DNA probes generated in LA-PCR. Computer analysis of the localization of the restriction sites in human genome has allowed us to propose an efficient technique for generating DNA probes by LA-PCR using the restriction endonucleases <i>Hae</i>III and <i>Rsa</i>I. In silico hydrolysis of human genomic DNA with endonucleases <i>Hae</i>III and <i>Rsa</i>I demonstrate that 100- to 1,000-bp DNA fragments are more abundant in the gene-rich regions. Applying in situ hybridization to metaphase chromosomes, we demonstrated that the produced DNA probes predominantly hybridized to the C-negative chromosomal regions, whereas the FISH signal was almost absent in the C-positive regions. The described protocol for generating DNA probes may be successfully used in subsequent cytogenetic analysis of the C-negative chromosomal regions.