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The correct insert was confirmed by PCR screening and nucleotide sequence confirmation (MWG Biotech, Germany).
Because of the stability and unique structure of PNAs, traditional sequence confirmation methods are not effective.
For further sequence confirmation, the fragments were sequenced using the M13 forward and gene-specific primers.
From this study, using the described methodology, NoV genomes were frequently detected in fresh produce however sequence confirmation was not successful for the majority of the samples tested.
In the present study, HPLC ESI IT (ion-trap) MS was used for carboxyterminal (C-terminal) amino acid sequence confirmation of intact recombinant Hirudin Variant 3 (HV3) and alkylated HV3.
After sequence confirmation, altered msn DNAs were sub-cloned into the pH Stinger vector.
After sequence confirmation of all four PCR products, one clone was selected for aggregation.
Purified plasmids were submitted to the UNMC core facility for sequence confirmation.
However, restriction sites can easily be incorporated into LIC tails to facilitate downstream sequence confirmation of the insert.
After sequence confirmation, fragments were recloned to murine stem cell virus (MSCV -based MSCV -basedor for retroviral expression.
Following transformation into E coli four colonies from each were picked for DNA sequence confirmation of the insert.
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