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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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Signal Pattern Problem in Real-time PCR
Posted by: cobanog (IP Hidden, New member, 1)
Date: December 19, 2005 01:38PM
Dear All;
We are using Opticon 2 Real time from MJ Research. For an assay, we use two molecular beacon probes which should hybridize with the amplified segments based on designed X and Y specific primer pairs from @#$%& chromosomes. Assay perfectly works with genomics DNA. Also, when we use embryo biopsies the signal intensity is high enough to surpass threshold line from Y chromosome. BUT we have certain pattern, which always starts from a high level and goes down to a low in a linear way from X chromosome. In addition, the signal pattern from amplified product based on X-specific primers was the same in our repeats (start with high and goes down to low level). As expected, we should have signal from X chromosome no matter either from male or female @#$%& embryos. Has anybody had an idea that why we could have such a signal pattern with X specific primers from X chromosome while it gives a signal pattern starting from low level and goes to high as number of cycle increase from Y chromosome as expected at successful PCR reaction? Thanks for your response in advance. Ozden
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