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Mutasyon Analiz Teknikleri I
Hafta VII PCR Temelli Genetik Analiz Yaklaşımları Mutasyon Analiz Teknikleri I Doç. Dr. Hilâl Özdağ
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DHPLC Transgenomic/WAVE
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DHPLC http://insertion.stanford.edu/melt.html
BRCA ex1 CAATAGGTAGCGATTCTGACCTTCGTACAGCAATTACTGTGATGCAATAA GCCGCAACTGGAAGAGTAGAGGCTAGAGGGCAGGCACTTTATGGCAAACT CAGGTAGAATTCTTCCTCTTCCGTCTCTTTCCTTTTACGTCATCCGGGGG CAGACTGGGTGGCCAATCCAGAGCCCCGAGAGACGCTTGGCTCTTTCTGT CCCTCCCATCCTCTGATTGTACCTTGATTTCGTATTCTGAGAGGCTGCTG CTTAGCGGTAGCCCCTTGGTTTCCGTGGCAACGGAAAAGCGCGGGAATTA CAGATAAATTAAAACTGCGACTGCGCGGCGTGAGCTCGCTGAGACTTCCT GGACGGGGGACAGGCTGTGGGGTTTCTCAGATAACTGGGCCCCTGCGCTC AGGAGGCCTTCACCCTCTGCTCTGGGTAAAGGTAGTAGAGTCCCGGGAAA GGGACAGGGGGCCCAAGTGATGCTCTGGGGTACTGGCGTGGGAGAGTGGA TTTCCGAAGCTGACAGATGGGTATTCTTTGACGGGGGGTAGGGGCGGAAC CTGAGAGGCGTAAGGCGTTGTGAACCCTGGGGAGGGGGGCAGTTTGTAGG TCGCGAGGGAAGCGCTGAGGATCAGGAAGGGGGCACTGAGTGTCCGTGGG GGAATCCTCGTGATAGGAACTGGAATATGCCTTGAGGGGGACACTATGTC TTTAAAAACGTCGGCTGGTCATGAGGTCAGGAGTTCCAGACCAGCCTGAC CAACGTGGTGAAACTCCGTCTCTACTAAAAATACAAAAATTAGCCGGGCG TGGTGCCGCTCCAGCTACTCAGGAGGCTGAGGCAGGAGAATCGCTAGAAC C with 851 base pairs Melting temperature of entire strand: 61.25° Temperature = 55° Celsius: 1-3, Temperature = 56° Celsius: 4-5, Temperature = 57° Celsius: , Temperature = 58° Celsius: 6-6, , , Temperature = 59° Celsius: 7-48, , , , Temperature = 60° Celsius: , , , , Temperature = 61° Celsius: , , , Temperature = 62° Celsius: , This sequence should be run at all of the following temperatures to ensure detection of all polymorphisms. Recommended temperature: 62° Recommended temperature: 56°
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DHPLC CBP Exon 6 A. Rothan’dan
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DHPLC A. Rothan’dan
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DHPLC A. Rothan’dan
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DHPLC Ezzeldin et al. Ann Biochem. 2002;306:63-73.
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DHPLC
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DHPLC DHPLC = denaturing high-performance liquid chromatography.
Ezzeldin et al. Ann Biochem. 2002;306:63-73.
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Karıştırma testi DHPLC
P300/EXON16 4239G->T E1014X
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Karıştırma testi DHPLC
CBP/EXON31.1 5371-8INSA
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DHPLC HDAC2 EXON1 Karıştırma
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HDAC2 143InsCAG
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DHPLC HDAC2 EXON12 Karıştırma
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HDAC2 1573DelA HCT15
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HSUV39H2 -11InsA (?) SSCP
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Kapiller Elektroforez
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Genel Primerlerle işaretleme
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P G>T (E1014X)
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P G>T (E1014X )
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CBP IVS21-4del22
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CBP IVS21-4del22
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CBP IVS31-7InsA
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CBP IVS31-7InsA
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TGCE Temperature Gradient Capillary Electrophoresis
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