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  • 1
    Electronic Resource
    Electronic Resource
    Estimating European admixture in African Americans by using microsatellites and a microsatellite haplotype (CD4/Alu) (1999)
    Springer
    Human genetics 〈Berlin〉 104 (1999), S. 149-157 
    Details
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have analyzed 10 unlinked microsatellites and a linked Alu deletion polymorphism at the CD4 locus in an African American population sample from Chicago (USA). Heterozygosity estimates at the microsatellite loci range from 0.727±0.025 (D3S1358) to 0.873±0.017 (D18S51), with an average of 0.794±0.016. These values are comparable to or higher than those reported for Europeans, with only one exception (D3S1358). The CD4/Alu haplotypic diversity (0.887±0.012) is comparable to diversity levels observed in sub-Saharan African populations and is higher than the diversity levels reported in European populations. No consistent pattern of within, between, or multi-locus deviations from Hardy-Weinberg expectations is observed, suggesting a low sub-heterogeneity within the sampled population. We have applied a maximum likelihood method and estimated the proportion of European admixture to the African American gene pool to be 0.26±0.02. The narrow confidence interval indicates that allele frequency data from multiple microsatellite loci, whether analyzed independently or as haplotypes, are particularly useful for estimating genetic admixture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004390050928
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  • 2
    Electronic Resource
    Electronic Resource
    Improved typing of human serum transferrin by isoelectric focusing on ultrathin layer polyacrylamide slab gels (1982)
    Springer
    Human genetics 〈Berlin〉 61 (1982), S. 39-41 
    Details
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An improved method for separating transferrin subtypes through the use of isoelectric focusing on ultrathin layer polyacrylamide gels is described. The most considerable problems encountered in the assessment of the Tf phenotypes after pretreatment with iron donor compounds are pointed out. Useful technical devices for reducing the occurrence of artifactitious extrabands are suggested. Finally, the gene frequencies in three samples from Italian populations are reported.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00291329
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  • 3
    Electronic Resource
    Electronic Resource
    Characterization of genetic variants of human serum transferrin by isoelectric focusing: Comparison between conventional and immobilized pH gradients, and application to a protocol for paternity testing (1988)
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988), S. 411-417 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The polymorphism of transferrin (Tf) is currently being studied by isoelectric focusing in carrier ampholyte-generated pH gradients, carrier ampholyte-separator pH gradients or in immobilized pH gradients. Details for obtaining reproducible results with each of the three procedures are outlined. The effectiveness of pretreatment of serum samples with ferrous/ferric salts is discussed, and incubation times optimized after spectrophotometric measurement of the monoferric Tf conversion. Most of the presently available commercial batches of carrier ampholytes do not reliably discriminate the six common TfC subtypes. Resolution of C1, C3 and C2 was achieved by adding 20 to 90 mM HEPES slab gels prepared with various carrier ampholytes. Isoelectric focusing in carrier ampholyte-separator pH gradients cannot be recommended as a standard typing procedure because the results strongly depend on the batch of carrier ampholytes. Tf subtype resolution was only achieved by using isoelectric focusing in immobilized pH gradients with pH slopes reliably reproducible from one experiment to another. Two major shortcomings of immobilized pH gradients are a marked tendency to protein precipitation at the application site and an interaction of proteins with the charged matrix. A protocol for Tf subtyping in immobilized pH gradients is described, based on prior desialylation of samples instead of pretreatment with iron. Sample entry into the matrix was optimized by addition of 5 mM Tris to the gels, and initially running them at low voltage. Recommendations are provided for the application of Tf typing for paternity testing.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150090811
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  • 4
    Electronic Resource
    Electronic Resource
    Simplified molding procedure for ultrathin-layer polyacrylamide slab gels using methacrylate supports and unsilanized glass plates: Hybrid isoelectric focusing on ultrathin immobilized pH gradients (1987)
    Weinheim : Wiley-Blackwell
    Electrophoresis 8 (1987), S. 371-373 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A simplified procedure for the preparation of ultrathin polyacrylamide slab gels is reported. The molding cells consist of an unsilanized glass plate and methacrylate plate or acryl-coated glass plate. Owing to the repelling properties of acrylate the ultrathin gels spontaneously detach from the acrylate surface on unfastening the molds, firmly adhering to the unsilanized glass. Gels as thin as 0.20-0.25 mm could be polymerized without resorting to silanized glass plates or polyester films. The procedure is especially suitable for sticky gels, such as highly cross-linked polyacrylamide gels and gels with immobilized pH gradients.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150080808
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  • 5
    Electronic Resource
    Electronic Resource
    Errors in sizing bands of hypervariable DNA profiles on autoradiograms: Are they Gaussian? (1992)
    Weinheim : Wiley-Blackwell
    Electrophoresis 13 (1992), S. 341-345 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The accuracy of procedures for sizing hypervariable restriction fragments by Southern blot analysis (SBA) has been tested under three different experimental conditions: (i) intrablot serial analyses: three heterozygous DNA profiles were tested 14 times each in the same gel electrophoresis; (ii) intralaboratory analyses: we replicated three profiles (six autoradiographic bands) in over 100 SBA experiments; (iii) interlaboratory analyses: 15 serial measurements produced in a recent collaborative study (Forensic Sci. Int. 1991, 49, 1-15) were taken into account. In these three cases, a typical U-shaped correlation curve between molecular size and coefficient of variation was found. We explain decrease of accuracy at both extremities of the gels in terms of: (i) enlarged shapes of bands and mean electrophoretic resolution at the cathode; (ii) diffusion and blurring of bands at the anodal edge. The three populations of data were subjected to a χ2 test and to the Kolmogorov-Smirnov test in-order to verify their compliance with normal distribution. Twenty-three out of 27 tests indicated no significant deviation from the assumption of Gaussian distribution. We recommend the adoption of tests for normality to validate the use of symmetric confidence intervals for calculating gene frequencies and asserting a match between adjacent bands.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150130171
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  • 6
    Electronic Resource
    Electronic Resource
    Improved classification of alpha-1-antitrypsin in immobilized pH gradients containing sucrose (1985)
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 402-404 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Two narrow immobilized pH gradients (pH 4.35-4.85 and pH 4.40-4.80) were formulated by linear interpolation from standard 1 pH unit ranges. Both gradients proved excellent for alpha-1-antitrypsin phenotype separation. Gel casting was simplified and the properties of the gels during the run improved by replacing glycerol with sucrose in the polymerization solutions.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150060810
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  • 7
    Electronic Resource
    Electronic Resource
    Immobilized pH gradients in ultrathin-layer (100 μm) gels: Avoiding capillarity by a ‘squeezing-sealing mold’ technique (1988)
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988), S. 514-519 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Several procedures for casting 100 μm ultrathin immobilized pH gradients are described. When acrylate/glass molding cassettes (Pascali et al., Electrophoresis 1987, 8, 371-373) are used two main problems are encountered: (i) a tendency of polymerization solutions, at the beginning of delivery, to spread across the glass surfaces with troublesome effects on the gradient stratification, and (ii) the raising of steep menisci at both extremities of the pH intervals, originating from capillarity phenomena and resulting in nonuniform gradients with bowed electrophoretic patterns. The first shortcoming was acceptably solved by increasing the density of sucrose gradients, and pouring them into prewarmed molding cassettes. The detrimental effect of menisci could be overcome by using a ‘squeezing-sealing mold’ technique. A molding cassette was endowed with a continuous, squared spacer frame, the upper side being open by inserting a wedged clip. A slight excess of polymerization solution was first dispensed into the cassette and squeezed away on removal of the clip. By completely excluding air from the molding cassette, uniform and well reproducible ultrathin gels could be cast. A major advantage of ultrathin immobilized pH gradient gels is the drastically shorter focusing time.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150090911
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  • 8
    Electronic Resource
    Electronic Resource
    The use of immobilized pH gradients for the detection of human polymorphisms in the forensic identification of bloodstains (1992)
    Weinheim : Wiley-Blackwell
    Electrophoresis 13 (1992), S. 239-243 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Some polymorphic proteins (α1-antitrypsin, orosomucoid, transferrin, group specific component, plasminogen) and enzymes (phosphoglucomutase, acid phosphatase, estrase D) were determined in bloodstain extracts by isoelectric focusing with carrier ampholytes (CA) and with immobilized pH gradients (IPGs) rehydrated with CA. IPGs yield superior results for typing of genetics markers in bloodstains since phenotypes are better distinguished and the bands are straighter and sharper in the presence of contaminants. Also, the sensitivity of IPGs with CA is similar to isoelectric focusing (IEF) with CA. A new variant, ACP*B1, found in Negroid west African populations and not found in Caucasians is described. Such a variant can only be determined by IPGs since its isoelectric point (pI 5.95) is close to that of the ACP*B (pI 6.05) variant.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150130149
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  • 9
    Electronic Resource
    Electronic Resource
    Efficient, small scale electroelution of high molecular weight DNA from agarose gels by a miniature vertical electrophoresis cell (1991)
    Weinheim : Wiley-Blackwell
    Electrophoresis 12 (1991), S. 317-320 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A miniature, vertical electrophoretic system is described that allows quick elution and efficient harvesting of double-stranded, high molecular weight DNA fragments from agarose gels. A gel slice containing the band of interest is excised from the agarose gel after submarine electrophoresis and placed into an electrophoresis cell, assembled by inserting a truncated micropipette tip into a 2.0 mL polypropylene tube. The tip supports the agarose slice and is connected to dialysis tubing. Platinum wire electrodes are placed over the tip and at the bottom of the test tube. DNA molecules are quickly eluted into the dialysis bag (60 s to 10 min for bands ranging from 2 to 23 kb) and then easily recovered. The method is highly efficient and allows considerable time saving. Of special interest is its applicability to small starting amounts of DNA (〈0.5 μg per band). Purity of the DNA is comparable, if higher, to that obtainable by conventional electroelution on dialysis bags and DEAE-cellulose membrane electrophoresis. The system is simple enough to be used as a routine method for eluting multiple bands in short times.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150120417
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  • 10
    Electronic Resource
    Electronic Resource
    Fluorescence-based classification of microsatellites using a single-wavelength semiautomatic sequencer: Genotype assignment and identity tests by analysis of comigrating peak profiles (1995)
    Weinheim : Wiley-Blackwell
    Electrophoresis 16 (1995), S. 1875-1880 
    Details
    ISSN: 0173-0835
    Keywords: Microsatellites ; Semiautomatic sequencers ; Identity tests ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Microsatellite analysis, based on fluorescein labeling and reading through a semiautomatic single wavelength sequencer, is described. Paris of labeled polymerase chain reaction (PCR) samples, mixed in equimolar proportion, were electrophoresed and the specific peaks read in a single gel lane. Identity was asserted when peaks overlapped in a unique fluorescent signal which, compared with individual sample profiles, had a twofold intensity. Classification was achieved by blending individual PCR products to ‘locus specific allelic ladders’ (composite samples containing a repertory of fragments allelic to a given locus) and by noticing the specific peak enhancement. The resulting protocol of analysis assigned no size and classified allelic forms by tandem repeat number. Applied to a large repertory of PCR products and compared with manual electrophoresis, this protocol proved to be reliable and reduced times and costs of genotype analysis. Analysis of comigrating peak profiles is highly objective and provides convincing evidence for diagnostics and identity tests.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.11501601307
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