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举报
2016-10-10 21:03
实时定量荧光PCR |
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举报
2016-10-07 03:57
It would be better that a single Western blot be used and show both 115 kDa target protein AND 37 kDa loading control in order to understand the shRNA experiment results truly. The shRNA lentivirus infection may be toxic to protein synthesis in infected cells, specifically by shRNA 2 and 3, while target mRNA knockdown by shRNA 1 shows effective, but RNA transcription is not inhibited around 72 hours postinfectionThe inhibition of protein synthesis by lentiviral vector infection could be recovered in shRNA 1 and in shRNA 2, but not in shRNA 3 ninety six hours postinfection, at this time point it reveals strong target mRNA knockdown by shRNA 1, moderated inhibition by shRNA 2, less effective inhibition by shRNA 3. A suggestion that a combined lentivirus infection with those producing shRNA 1, and shRNA 3 and shRNA 2 be done to find best shRNA combination showing effective target mRNA knockdown while posing no harm to cellular protein synthesis. |
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