I cloned a 5Kb gene and linked into a common T vector which is about 3Kb. After screen with Amp i take it with normal PCR but the result of PCR is weired, there are two specific bands one is 3Kb more bright and another is 5Kb alittle weak. If the 3Kb band is the vector jioned a primer dimers, but how to illustrate the 5kb band?
Yuri Tokarev
my plasmids and amplicon after GeneClean often produce two bands, most likely due to supercoiling of a portion of molecules. The electrophoretic mobility is different and two bands are visible, or single bands with different mobility in neighbor samples where single product of uniform size is expected.
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Liu Yuan wu
Thanks for your reply, after sequencing the 5kb band just a delay of 3kb fragment
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