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The resulting PCR products (528 bp) were subcloned using the TOPO TA Cloning Kit (Invitrogen, Darmstadt, Germany).
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The expected PCR fragment (700 bp) was subcloned using TOPO TA cloning kit (Invitrogen, CA) following the manufacturer's instructions.
The plasmids were subcloned using the Stratgene pPCRScriptamp kit.
All constructs were subcloned using the pCfvtx vector as described previously [6].
The resulting 705 bp product was subcloned using the TA-cloning vector pCR3.1 (Invitrogen, Paisley, UK) generating pCR-EPO.
A 2.6 kb PCR product containing 256 bp of flanking sequence was subcloned using the pGemT-Easy vector (Promega).
PCR product was subcloned using TOPO TA Cloning Kit pCR®2.1 (Invitrogen) and 20 colonies per sample were sequenced.
In order to transfer the full gene, the amplified fragment of 615 bp was subcloned into a high expression vector, pYES2.
In this case, a truncated fragment of 1632 bp was subcloned, in which the N-terminal extension was removed in order to prevent incorrect folding of the recombinant protein.
The PHR region of gwCry1 (1536 bp) was subcloned into the polylinker region via NcoI/NotI digest.
The full length sequence of gwCry1a (1866 bp) was subcloned into the polylinker region of the pDIA92B-His transfervector by SmaI/NotI digest.
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