TY - DATA T1 - Supplementary Material for: A Method for Generating Selective DNA Probes for the Analysis of C-Negative Regions in Human Chromosomes PY - 2011/07/28 AU - Morozkin E.S. AU - Loseva E.M. AU - Karamysheva T.V. AU - Babenko V.N. AU - Laktionov P.P. AU - Vlassov V.V. AU - Rubtsov N.B. UR - 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 DO - 10.6084/m9.figshare.5122642.v1 L4 - https://ndownloader.figshare.com/files/8707714 KW - C-negative regions KW - Linker-adapter polymerase chain reaction (LA-PCR) KW - FISH KW - Supernumerary marker chromosomes N2 - 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 HaeIII and RsaI. In silico hydrolysis of human genomic DNA with endonucleases HaeIII and RsaI 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. ER -