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การพัฒนาการตรวจสอบโครโมโซมเพศของมนุษย์ด้วยเทคนิค Fluorescent in situ Hybridization และ Real time PCR (Development of Human Sex Chromosome Detection for Fluorescent in situ Hybridization and Real Time PCR Techniques) |
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| รหัสดีโอไอ | |
| Creator | สุพัตรา พรวิลาศสิริ |
| Title | การพัฒนาการตรวจสอบโครโมโซมเพศของมนุษย์ด้วยเทคนิค Fluorescent in situ Hybridization และ Real time PCR (Development of Human Sex Chromosome Detection for Fluorescent in situ Hybridization and Real Time PCR Techniques) |
| Publisher | Genetics Society of Thailand |
| Publication Year | 2556 |
| Journal Title | Thai Journal of Genetics |
| Journal Vol. | S.1 |
| Journal No. | S1 (Special Issue 1) |
| Page no. | 179 |
| Keyword | fluorescent in situ hybridization, real time PCR, sex chromosome |
| ISSN | 8578664 |
| Abstract | Nowsadays fluorescent in situ hybridization (FISH) and polymerase chain reaction (PCR) techniques play role in genetic material identification, genotyping, gene mutation detection and genetic disorder diagnosis etc. PCR technique, especially, real time PCR (RT PCR) is a technique which gives rapid and high precisely results. This research, we develop the detection of DXZ and SRY for FISH and RT PCR. DXZ and SRY probes for FISH have the size of 580 bp and 470 bp, respectively. Both probes are hybridized on human male lymphocytes and found 80% of lymphocytes showing both probe signals. For RT PCR, the amplified fragment of DXZ and SRY are 184 bp and 214 bp, respectively. The detection is used SYBR green dye and 147 bp of human actin is used as the control. The effectiveness of DXZ, SRY and human actin are 89.2, 52 and 68, while the correlation coefficience are 0.994, 0.949 and 0.935, respectively. These results show the efficiency of both developed techniques for human sex chromosome diagnosis and crossing over between X and Y chromosome detection. We are still improving both methods to get more efficiency for single cell detection. This will be fruitful for decreasing import of detection kit from abroad. |
