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數(shù)峰青1989鐵蟲 (小有名氣)
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[求助]
5'race 沒有結(jié)果,求高人指點
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近一段時間在用takara的試劑盒做5'race,但是第二次PCR之后怎么也得不到一個清晰的條帶。。。實驗結(jié)果如所附圖,請高人指點一二 RNA分離結(jié)果(該結(jié)果說明rna提取很成功,可以做下一步實驗).jpg 2次PCR之后產(chǎn)物電泳結(jié)果(沒有條帶,原因是什么,請高人指點).jpg |
木蟲 (著名寫手)
鐵蟲 (小有名氣)
| 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. |

鐵蟲 (小有名氣)
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