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Linkage Analysis Of Myp12 And Myp14 In Families From Lahore

By: Maryam Zahra | Prof.Dr.Masroor Elahi Babar.
Contributor(s): Dr. Ali Raza Awan | Prof. Dr.
Material type: materialTypeLabelBookPublisher: 2010Subject(s): Institute of Biochemistry & BiotechnologyDDC classification: 1216,T Dissertation note: Myopia is one of the most common refractive errors of the eye worldwide that can effect clarity of vision, limit occupational choices and contribute to increased risk to vision threatening conditions. Six families of different casts were enrolled from Lahore (Punjab). Total of six autosomal dominant families were screened for linkage to the known nonsyndromic autosomal dominant and QTL myopia locus, MYP12 and MYP14 respectively. 5mL blood sample were collected aseptically in a 5Oml falcon tubes containing EDTA. DNA extraction was done by inorganic method. Three markers for each locus were selected from literature and redesigned by using 'primer 3' software. These markers were optimized for their annealing temperature and specific concentration of PCR ingredients by gradient PCR. After that all of the markers were amplified separately on genomic DNA samples of each family. PCR products of each of the marker were run on 1.2 % agarose gel along with 50 base pair ladder to visualize the bands of amplified products at 110 volts for 30 minutes. Linkage analyses were carried out by genotyping through PAGE unit of Major Science, model no. MV-2ODSYS. PCR products were run on Polyacrylamide Gel Electrophoresis (PAGE) to examine amplified bands of microsatellites. A standard 5Obp DNA ladder was run along with the sample PCR products as a reference. By reading the alleles appeared on gel haplotypes were constructed for each member of these families. The overall results of this study did not show evidence for linkage of myopia in thee families to the selected loci MYP12 and MYP14 on chromosomes 2 and 1 respectively. It might be possible any other identified locus or any new locus involved in this population of Pakistan. The findings represented here do not represent the conclusion of this study but do provide ongoing data for further investigation into the exact gentic causes of mypia in Pakistan.
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Veterinary Science 1216,T (Browse shelf) Available 1216,T
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Myopia is one of the most common refractive errors of the eye worldwide that can effect clarity of vision, limit occupational choices and contribute to increased risk to vision threatening conditions. Six families of different casts were enrolled from Lahore (Punjab). Total of six autosomal dominant families were screened for linkage to the known nonsyndromic autosomal dominant and QTL myopia locus, MYP12 and MYP14 respectively. 5mL blood sample were collected aseptically in a 5Oml falcon tubes containing EDTA. DNA extraction was done by inorganic method. Three markers for each locus were selected from literature and redesigned by using 'primer 3' software. These markers were optimized for their annealing temperature and specific concentration of PCR ingredients by gradient PCR. After that all of the markers were amplified separately on genomic DNA samples of each family. PCR products of each of the marker were run on 1.2 % agarose gel along with 50 base pair ladder to visualize the bands of amplified products at 110 volts for 30 minutes. Linkage analyses were carried out by genotyping through PAGE unit of Major Science, model no. MV-2ODSYS. PCR products were run on Polyacrylamide Gel Electrophoresis (PAGE) to examine amplified bands of microsatellites. A standard 5Obp DNA ladder was run along with the sample PCR products as a reference. By reading the alleles appeared on gel haplotypes were constructed for each member of these families. The overall results of this study did not show evidence for linkage of myopia in thee families to the selected loci MYP12 and MYP14 on chromosomes 2 and 1 respectively. It might be possible any other identified locus or any new locus involved in this population of Pakistan. The findings represented here do not represent the conclusion of this study but do provide ongoing data for further investigation into the exact gentic causes of mypia in Pakistan.

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