Concentrations and Absorbance Ratios for Samples Isolated with the MasterPure Genomic DNA Purification Kit. 1691 results in a GtoA base substitution. The Factor V gene status of the patient can subsequently be determined by direct visualization of digestion fragments after agarose gel electrophoresis. The genomic DNA obtained was amplified using primers specific for a 267 base-pair region spanning nucleotide position 1691 in the Factor V gene. DNA from a homozygous FVR506Q patient would result in fragments of 200 and 67 base pairs.
Heterozygous individuals show a combination of the two patterns.
All samples in Figure 1 show the presence of the 163 bp fragment and the absence of the 200 bp fragment, indicating these DNA samples were derived from homozygous normal individuals. Fragment sizes are indicated. The purified genomic DNA was also used to detect mutations associated with Fragile X syndrome by Southern transfer and hybridization. Southern blot analysis of digested genomic DNA was used to assess the size and degree of methylation of the CGG repeat region of the FMR 1 gene. I is methylation sensitive and thus will not cut when its recognition sequence has been methylated, as in association with the Fragile X disease or as a normal process of X-chromosome inactivation in females.