pELMO and pGEM-T Easy cloning efficiency regarding low, medium and large sized inserts.
For easy cloning of any antibody gene without potential modification of the antibody sequence, restriction sites were introduced in the pelB sequence and following the termination codon.
Plasmid identity was confirmed by sequencing, using the ccdBsec-F/ccdBsec-R and SP6/T7 primer sets; these were then aligned with the corresponding pELMO and pGEM-T Easy cloning site flanking sequences.
Every Gateway vector possesses att sites necessary for recombination and easy cloning of cDNA.
PCR products were cloned using the pGEM-T Easy cloning system (Clontech) and sequenced with Applied Biosystems Big dye terminator kit v1.1 and the ABCB7 forward primer.
The amplified fragment was cloned into pGEM-T easy cloning vector (Promega), sequenced.
Products were ligated into pGEM-T Easy Cloning Vector (Promega, Southampton, United Kingdom).
The amplified silencing fragments were cloned into the pGEM-T vector (pGEM-T Easy cloning kit; Promega, UK).
The amplified sequences were cloned into pGEM-T easy cloning system (Promega) and transformed into Escherichia coli DH5α cells.
PCR products were cloned into pGEM-T Easy cloning vector by using the pGEM-T Easy Vector System I Kit (Promega), in accordance with the manufacturer's instructions.
The PCR products were cloned with the pGEM®-T Easy cloning kit (Promega USA) and XL1 Blue Escherichia coli competent cells.
