Mutasyon Analiz Teknikleri I Hafta VII PCR Temelli Genetik Analiz Yaklaşımları Mutasyon Analiz Teknikleri I Doç. Dr. Hilâl Özdağ

DHPLC Transgenomic/WAVE http://www.transgenomic.com/flash/DHPLCFlash2.asp

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: 777-789, Temperature = 58° Celsius: 6-6,776-776,790-791, Temperature = 59° Celsius: 7-48,302-312,773-775,792-792, Temperature = 60° Celsius: 49-138,297-301,313-314,771-772, Temperature = 61° Celsius: 139-296,315-316,654-770,793-851 Temperature = 62° Celsius: 317-653, This sequence should be run at all of the following temperatures to ensure detection of all polymorphisms. Recommended temperature: 62° Recommended temperature: 56° http://insertion.stanford.edu/melt.html

DHPLC CBP Exon 6 A. Rothan’dan

DHPLC A. Rothan’dan

DHPLC A. Rothan’dan

DHPLC Ezzeldin et al. Ann Biochem. 2002;306:63-73.

DHPLC

DHPLC DHPLC = denaturing high-performance liquid chromatography. Ezzeldin et al. Ann Biochem. 2002;306:63-73.

Karıştırma testi DHPLC P300/EXON16 4239G->T E1014X

Karıştırma testi DHPLC CBP/EXON31.1 5371-8INSA

DHPLC HDAC2 EXON1 Karıştırma

HDAC2 143InsCAG

DHPLC HDAC2 EXON12 Karıştırma

HDAC2 1573DelA HCT15

HSUV39H2 -11InsA (?) SSCP

Kapiller Elektroforez

Genel Primerlerle işaretleme

P300 4239G>T (E1014X)

P300 4239G>T (E1014X )

CBP IVS21-4del22

CBP IVS21-4del22

CBP IVS31-7InsA

CBP IVS31-7InsA

TGCE Temperature Gradient Capillary Electrophoresis http://www.spectrumedix.com/7144_9155163437/blank/browse.asp?a=383&BMDRN=2000&BCOB=0&c=53494