I am trying to amplify my gene of interest from my recombinant bacmid isolated from Dh10Bac cells using gene-specific primers. However, the PCR is giving multiple non-specific bands in the amplification as visualized on gel. However, the positive control is giving a single correct sized PCR product. Thus, I would like to validate my primer pair specificity against the native baculovirus shuttle vector (pMON14272) and the helper plasmid (pMON7142) and check whether they allow for non-specific product amplification due to mispriming. I have tried searching for the sequences online but have been so far unable to find them. Any help in this regard would be much appreciated.
Hi Shiv,
did you ever find an answer to this? I'm after the same information.
thanks
Unfortunately, I could not find such data, however, there are some bacmid sequences published by Geneva Biotech for their Multibac systems.
PFA link to the site: https://geneva-biotech.com/product_category/insect-cell-expression/multibac/vector-sequences/
The sequences for the bacmid: DH10MultiBac should be related to the original bacmid pMON14272
Thanks for the update, I have pieced together partial sequences and maps, it was hard work but it all makes sense now!
Here is the full plasmid sequence suing long read seqeuncing. I just submitted it to genbank too.
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