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Biomedical subjects

B Budowle

Publications and source records attributed to B Budowle.

At least 145 records · Page 8Linked to original sources

Arab population data on the PCR-based loci: HLA-DQA1, LDLR, GYPA, HBGG, D7S8, Gc, and D1S80.

Allele and genotype frequencies for seven polymerase chain reaction (PCR)-based DNA genetic markers were determined in an Arab sample population. The loci analyzed were HLA-DQA1, LDLR, GYPA, HBGG, D7S8, Gc and D1S80. Results were obtained from the first six loci using the AmpliType HLA-DQ alpha DNA and AmpliType PM PCR Amplification and Typing Kits. The VNTR locus D1S80 PCR product was analyzed by polyacrylamide electrophoresis and silver staining. All loci meet Hardy-Weinberg expectations. The frequency data can be used in forensic analyses and paternity tests to estimate the frequency of a DNA profile in the Arab population.

Alleles↗

Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts.

Techniques have been developed for extracting, amplifying and directly sequencing mitochondrial DNA (mtDNA) from human hair shafts. The hair shaft is ground in a glass micro-tissue grinder, and the DNA is extracted with organic solvent and purified by filtration. The filtrate subsequently provides the mtDNA template for the PCR. The two hypervariable segments of the mtDNA control region are amplified in four separate reactions. After a purification step to remove unincorporated PCR primers, amplified products are quantitated by capillary electrophoresis and subjected to cycle sequencing. The products are separated and analyzed on an automated DNA sequencer. The mtDNA sequences from the hair shaft match the mtDNA sequences from blood samples taken from the same donor.

Base Sequence↗

Fixed bin frequency distributions for the VNTR locus D5S110 in general United States reference databases.

Fixed bin frequencies for the D5S110 locus were determined in African Americans, Caucasians, Southeastern Hispanics, and Southwestern Hispanics. The data were generated by RFLP analysis of HAE III-digested genomic DNA. The D5S110 locus met Hardy-Weinberg expectations in the four sample populations, and there is no evidence for association of alleles between the D5S110 and other routinely used VNTR loci. The frequency data can be used in forensic analyses and paternity tests to estimate the frequency of a DNA profile in various general United States populations.

Black People↗

Fixed bin population data for the VNTR loci D1S7, D2S44, D4S139, D5S110, and D17S79 in Chinese from Taiwan.

Fixed bin frequencies for the VNTR loci D1S7, D2S44, D4S139, D5S110, and D17S79 were determined in a Chinese sample population. The data were generated by RFLP analysis of Hae III-digested genomic DNA and chemiluminescence detection. The five VNTR loci meet Hardy-Weinberg expectations in the Chinese sample population, and there is little evidence for association of alleles between the VNTR loci. The frequency data can be used in forensic analyses and paternity tests to estimate the frequency of a DNA profile in Chinese.

Asian People↗

D1S80 population data in African Americans, Caucasians, southeastern Hispanics, southwestern Hispanics, and Orientals.

Allele frequencies for the locus D1S80 were determined in African American, Caucasian, Southeastern Hispanic, Southwestern Hispanic, and Oriental sample populations using the polymerase chain reaction and subsequent electrophoresis and silver staining of the amplified products. Due to the presence of anodal and cathodal electrophoretic variants (in reference to the steps in an allelic ladder), allele frequencies were established using a classification protocol based on the steps in the allelic ladder. All sample populations met Hardy-Weinberg expectations for D1S80. In addition, there was no evidence for association of alleles between the loci D1S80 and D1S7. The product of allele frequencies from the data from the sample populations in this study can be used in forensic analyses and paternity tests to estimate the frequency of a D1S80 DNA genotype.

Alleles↗

Validation and population studies of the loci LDLR, GYPA, HBGG, D7S8, and Gc (PM loci), and HLA-DQ alpha using a multiplex amplification and typing procedure.

Studies were performed to evaluate the forensic applicability of multiplex amplification of the loci low density lipoprotein receptor, glycophorin A, hemoglobin G gammaglobin, D7S8, and group-specific component (PM loci) and simultaneous typing of these loci using a reverse dot blot approach where allele specific oligonucleotide probes are immobilized on a nylon membrane strip. These results were obtained by using the AmpliType PM PCR Amplification and Typing Kit. The experiments included: mixed body fluid studies; chemical contaminant effects on the DNA in body fluid samples; the effect of typing DNA from body fluid samples deposited on various substrates; the effect of microorganism contamination on typing DNA derived from blood and semen; the effect of sunlight and storage conditions on DNA typing; determination of the sensitivity of detection of the PM test kit; determination of cross-reactivity of DNA from species other than human; typing DNA derived from various tissues from an individual; and an evaluation of the hybridization temperature of the assay. The data demonstrate that DNA exposed to a variety of environmental insults yields reliable PM typing results. Allele and genotype frequencies for six loci (PM loci and HLA-DQ alpha) were determined in African Americans. Caucasians, southeastern Hispanics, and southwestern Hispanics. All loci meet Hardy-Weinberg expectations and there is little evidence for association of alleles between the loci. The frequency data can be used in forensic analyses and paternity tests to estimate the frequency of a multiple locus DNA profile in various general United States populations.

Alleles↗

A chemiluminescence-based detection system for human DNA quantitation and restriction fragment length polymorphism (RFLP) analysis.

A protocol for the chemiluminescent detection of restriction fragment length polymorphism (RFLP) profiles with the sensitivity of radioisotope-based detection systems is presented. RFLP profiles for the loci D2S44, D17S79, D1S7, D4S139, and D5S110 were obtained from ten nanograms of HaeIII-digested K562 DNA and 50 nanograms of human genomic DNA isolated from bloodstains. DNA is transferred to an amphoteric membrane using a neutral, high salt transfer procedure. Oligonucleotide probes directly conjugated to alkaline phosphatase are hybridized to the membranes, then the membranes are incubated with a substrate that emits light upon chemical cleavage by the alkaline phosphatase. Several variables were investigated to determine the optimum conditions for RFLP profile generation, among them transfer conditions, film selection, and exposure conditions. A chemiluminscent human DNA quantitation protocol, consistent with the RFLP detection protocol, is also presented.

Cross-Linking Reagents↗

A microsatellite polymorphism in the von Willebrand factor gene: comparison of allele frequencies in different population samples and evaluation for forensic medicine.

The allele frequencies at the tetranucleotide repeat (TCTA) vWA locus in the vWF gene were determined in the general Finnish population, in a population representing an internal isolate of Finland, in the Vologda-Russian population, and in US Black population samples. The allele and genotype frequencies from these population samples were compared with each other and with those reported from Spanish and British population samples. Statistically significant differences were demonstrated between most of the different groups (Finns vs. Vologda-Russians, Finns vs. US Blacks, Finns vs. Spanish, Vologda-Russians vs. US Blacks, Vologda-Russians vs. Spanish, US Blacks vs. Spanish and US Blacks vs. British Caucasians), but not between the two Caucasoid population samples from Finland and Great Britain, nor between or within the subpopulation samples from Finland and those from Vologda-Russia. In addition, the vWA marker was evaluated and demonstrated to be reliable for forensic purposes and paternity testing.

Adult↗

Quantitation of polymerase chain reaction products by capillary electrophoresis using laser fluorescence.

In samples where the amount of DNA is limited, the polymerase chain reaction (PCR) can amplify specific regions of the DNA. A quantitative analysis of the PCR product would be desirable to ensure sufficient DNA is available for analysis. In this study, we examine the use of capillary electrophoresis (CE) with laser fluorescence detection for quantitation of PCR products. A coated open tubular capillary was used with a non-gel sieving buffer and a fluorescent intercalating dye to obtain results within 20 minutes. Using an internal standard, peak migration time was below 0.1% relative standard deviation (R.S.D.) with a peak area precision of 3% R.S.D. In comparison to quantitation by hybridization, (i.e., slot blot) and spectrophotometric analysis, capillary electrophoresis shows distinct advantages due to its ability to separate unincorporated primers and PCR byproducts from the targeted PCR product. The results demonstrate that CE can be used to monitor the quality and quantity of the PCR product.

DNA, Viral↗

Swiss population data on the loci HLA-DQ alpha, LDLR, GYPA, HBGG, D7S8, Gc and D1S80.

Allele and genotype frequencies for seven polymerase chain reaction-based DNA genetic markers were determined in a Swiss sample population. The loci are D1S80, HLA-DQ alpha, low density lipoprotein receptor, glycophorin A, hemoglobin G gammaglobin, D7S8, and group-specific component. All loci meet Hardy-Weinberg expectations. In addition, there is no detectable association of alleles between loci. The frequency data can be used in forensic analyses and paternity tests to estimate the frequency of a DNA profile in the Swiss population.

DNA↗

Analysis of short tandem repeat (STR) HUMVWA in the Spanish population.

Amplification by polymerase chain reaction (PCR) of variable number of tandem repeat (VNTR) loci and subsequent typing by electrophoresis and silver staining has become a useful tool for identity testing. One viable group of genetic markers amenable to amplification by PCR is the short tandem repeat (STR) loci. A horizontal discontinuous polyacrylamide gel electrophoresis (PAGE) method was used to type the amplified products of the STR HUMVWA. Typing for VWA of 120 unrelated Spanish Caucasians was done. Six alleles were observed with frequencies in the range 0.096-0.242. The genotype distribution meets Hardy-Weinberg expectations (0.25 < P < 0.50). The heterozygosity was 73.3% and the discrimination power (DP) 0.94. Simultaneously, in a small sample of families (n = 24) no new mutations could be found.

Alleles↗

Horizontal polyacrylamide gel electrophoresis for the separation of DNA fragments.

A discontinuous borate/formate buffer system is presented for horizontal polyacrylamide gel electrophoresis of DNA fragments. The resolution potential of the system could be altered by changing the total monomer concentration (5-9%T), the concentration of the crosslinker piperazine diacrylamide (2-5%CPDA), as well as the concentration of formate in the gel (40-120 mM), the leading ion of the buffer system. The separation of DNA fragments would be improved by increasing the migration distance from 22 to 28 cm. This discontinuous polyacrylamide gel electrophoresis system proved highly reproducible.

Buffers↗

D1S80 allele frequencies in a Chinese population.

Allele frequencies for the VNTR locus D1S80 were determined in a Chinese population sample using the polymerase chain reaction and subsequent analysis of the amplified products by polyacrylamide gel electrophoresis and silver staining. A total of 18 nominal D1S80 alleles were observed in 105 unrelated Chinese. The data demonstrate that D1S80 is highly polymorphic in Chinese with a heterozygosity of 90.5%. The D1S80 frequency distribution meets Hardy-Weinberg expectations. This D1S80 data can be used in forensic analyses and paternity tests to estimate the frequency of a DNA profile in a Chinese population.

Alleles↗

Sequential multiplex amplification (SMA) of genetic loci: a method for recovering template DNA for subsequent analyses of additional loci.

A method called Sequential Multiplex Amplification (SMA) has been developed whereby a limited amount of DNA extracted from a sample can be reutilized for several single polymerase chain reaction (PCR) amplifications. The method involves recovery of genomic template DNA by microfiltration of PCR-amplified samples. Up to 5 different loci have been typed, each in a single system PCR-based assay, beginning with a test quantity of 5 ng template DNA. Genotypes of the DNA donors were compared with those obtained from individual amplifications and shown to be identical. This could be a useful technique for typing a number of loci from a limited amount of DNa and to recover template DNA from samples previously subjected to PCR. Obviously, when small quantities of template DNA are available, this technique can prove quite useful.

Base Sequence↗

Swiss population data on three tetrameric short tandem repeat loci--VWA, HUMTHO1, and F13A1--derived using multiplex PCR and laser fluorescence detection.

Allele and genotype frequencies for 3 tetrameric short tandem repeat loci VWA, HUMTHO1, and F13A1 were determined in a Swiss population sample using multiplex PCR and subsequent electrophoresis in DNA sequencing gels processed by automated laser fluorescence detection. The technique allows single base pair resolution and rapid typing, with a concomitant reduction in the potential for human transcriptional typing errors. All loci meet Hardy-Weinberg expectations. In addition, there is little evidence for association of alleles among the 3 loci. The allelic frequency data can be used in forensic analyses and paternity tests to estimate the frequency of a multiple STR locus DNA profile in the Swiss population.

Alleles↗

Greater differences in forensic DNA profile frequencies estimated from racial groups than from ethnic subgroups.

A National Research Council (NRC) report asserted that genetic diversity between subgroups within a race is greater than between races. While this is counterintuitive to the basic tenets of human population genetics, the forensic application of statistical estimates of DNA profile frequencies has been questioned. The general approach used for estimating DNA profile frequencies by the majority of North American forensic laboratories is the use of the product rule on fixed bin allele frequencies derived from general population group databases. An informal analysis of the differences in frequency estimates within Hinf I-generated RFLP data determined in three ethnically distinct population groups--Norwegians, Spanish, and Turks--and in Caucasians and Blacks from the United States is presented. The VNTR loci analyzed are D2S44, D7S21, and D12S11. The data demonstrate that the assertion by the NRC is unfounded and that major population group databases, in lieu of subpopulation databases, can be used to provide estimates of DNA profile frequencies without consequences of wrongful bias.

Black People↗

Identification of host DNA by amplified fragment length polymorphism analysis: preliminary analysis of human crab louse (Anoplura: Pediculidae) excreta.

The ability to identify individual hosts of hematophagous arthropods via bloodmeal analyses is a continuing pursuit in both medical and forensic entomology. Characterization of human DNA from blood-feeding arthropods has been advanced substantially by preparation techniques, such as the polymerase chain reaction (PCR). Successful application of amplified fragment length polymorphism (AMP-FLP) analysis to excreta obtained from adult crab lice, Pthirus pubis (L.), fed on human volunteers is reported herein. Human DNA derived from crab louse excreta was typed successfully for two human DNA genetic markers, D1S80 and HUMTH01. Although preliminary, these results illustrate the ability of AMP-FLP analyses to provide individual human locus characterizations from desiccated arthropod excrement.

Animals↗