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3Â¥: Originally posted by robustli at 2013-03-27 02:02:13
there are so many reasons such as the quality of the cDNA library, annealing temperature, primer specificity,elongation time. I guess you may need check them one by one. I usually use primer 5 to des ...

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Сµ¤Ä¾Ä¾: ½ð±Ò+2, ¹ÄÀøÓ¦Öú 2013-03-29 16:03:31
gyesang: »ØÌûÖö¥ 2013-05-31 16:15:43
there are so many reasons such as the quality of the cDNA library, annealing temperature, primer specificity,elongation time. I guess you may need check them one by one. I usually use primer 5 to design my primer, there are other primers design software you can choose. after this try different annealing temperature, sometimes you can add DMSO if the template is GC-rich. At the same time, give it enough extention time. If your target gene is rare, you can also try the nested PCR.
immunologyparasite
3Â¥2013-03-27 02:02:13
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