Mutagenized HLA DNA Constructs: Tools for Validating Molecular HLA Typing Methodologies

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Description

This study describes the development and validation of mutagenized cloned DNA constructs, which correspond to the polymorphic regions of the class II region of the HLA complex. The constructs were used to verify the allelic specificity of primers and probes in polymerase chain reaction (PCR)-based HLA typing assays such as Sequence Specific Primers (SSP) and Sequence Specific Oligonucleotide Probes (SSOP). The constructs consisted of the entire polymorphic region of exon 2 of class II HLA allele sequences that included primer annealing sites or probe hybridization sites. An HLA allele sequence was inserted into a plasmid, cloned, then mutagenized to match ... continued below

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viii, 154 leaves : ill. (some col.)

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Schulte, Kathleen Q. May 1999.

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  • Schulte, Kathleen Q.

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Description

This study describes the development and validation of mutagenized cloned DNA constructs, which correspond to the polymorphic regions of the class II region of the HLA complex. The constructs were used to verify the allelic specificity of primers and probes in polymerase chain reaction (PCR)-based HLA typing assays such as Sequence Specific Primers (SSP) and Sequence Specific Oligonucleotide Probes (SSOP). The constructs consisted of the entire polymorphic region of exon 2 of class II HLA allele sequences that included primer annealing sites or probe hybridization sites. An HLA allele sequence was inserted into a plasmid, cloned, then mutagenized to match a specific HLA allele, and finally, the correct clone was verified by bidirectional sequencing of the insert. Thus, the construct created a cloned reference DNA sample for any specific allele, and can be used to validate the accuracy of various molecular methodologies.

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viii, 154 leaves : ill. (some col.)

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  • May 1999

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  • March 9, 2015, 8:15 a.m.

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  • April 13, 2016, 3:49 p.m.

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Schulte, Kathleen Q. Mutagenized HLA DNA Constructs: Tools for Validating Molecular HLA Typing Methodologies, thesis, May 1999; Denton, Texas. (digital.library.unt.edu/ark:/67531/metadc500888/: accessed November 21, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; .