Linked Life Data

9144935

Source:http://linkedlifedata.com/resource/pubmed/id/9144935

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1997-6-5
pubmed:abstractText
An automated DNA sequencer utilizing high sensitivity infrared (IR) fluorescence technology together with Polymerase Chain Reaction (PCR) methodology was used to detect several sex differentiating loci on the X and Y chromosomes from various samples often encountered in forensic case work. Amplifications of the X-Y homologous amelogenin gene, the alpha-satellite (alphoid) repeat sequences and the X and Y chromosome zinc finger protein genes ZFX and ZFY (ZFX/ZFY) were performed. DNA extracted from various forensic specimens was amplified using either Taq, Tth or ThermoSequenase. Multiplexing using primers for all three loci in one reaction tube was achieved using Tth and ThermoSequenase. Two IR labeling strategies for detection of PCR products were utilized. In the first strategy, one of the PCR primers contained a 19-base extension at its 5' end identical to an IR-labeled universal M13 Forward (-29) primer which was included in the amplification reactions. During PCR the tailed primer generates sequence complementary to the M13 primer which subsequently primes the initial amplification products, thereby generating IR-labeled PCR products. In the second strategy, dATP labeled with an IR dye (IR-dATP) was included in the amplification reaction. During amplification IR-dATP was utilized by the polymerase and incorporated into the synthesized DNA, thus resulting in IR-labeled PCR products. X and Y specific bands were readily detected using both labeling methodologies. Amplified products were electrophoretically resolved using denaturing Long-Ranger gels and detected with an automated detection system using IR laser irradiation. A separation distance of 15 cm allowed run times of less than 2 h from sample loading to detection. Because the gels could be run more than once, at least 120 samples (2 loads x 60 samples/load) can be typed using a single gel.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0022-1198
pubmed:author
pubmed:issnType
Print
pubmed:volume
42
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
452-60
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed-meshheading:9144935-Alleles, pubmed-meshheading:9144935-Amelogenin, pubmed-meshheading:9144935-DNA, Satellite, pubmed-meshheading:9144935-DNA-Binding Proteins, pubmed-meshheading:9144935-Dental Enamel Proteins, pubmed-meshheading:9144935-Female, pubmed-meshheading:9144935-Fluorescent Dyes, pubmed-meshheading:9144935-Humans, pubmed-meshheading:9144935-Kruppel-Like Transcription Factors, pubmed-meshheading:9144935-Male, pubmed-meshheading:9144935-Polymerase Chain Reaction, pubmed-meshheading:9144935-Repetitive Sequences, Nucleic Acid, pubmed-meshheading:9144935-Sex Determination Analysis, pubmed-meshheading:9144935-Spectrophotometry, Infrared, pubmed-meshheading:9144935-Tooth Germ, pubmed-meshheading:9144935-Transcription Factors, pubmed-meshheading:9144935-X Chromosome, pubmed-meshheading:9144935-Y Chromosome, pubmed-meshheading:9144935-Zinc Fingers
pubmed:year
1997
pubmed:articleTitle
Infrared fluorescent detection of PCR amplified gender identifying alleles.
pubmed:affiliation
Nebraska State Patrol Criminalistics Laboratory, Lincoln 68502, USA.
pubmed:publicationType
Journal Article