At Addgene, we sequence all incoming plasmids using Next Generation Sequencing (NGS). We use plexWell library preparation technology provided by seqWell coupled with Illumina sequencing to sequence and assemble full plasmid sequences. Addgene then uses SnapGene software to analyze the NGS results to validate the identity of important plasmid elements including the insert, essential backbone elements, any mutations indicated during the deposit process (ex: mismatches, truncations, insertions, etc), and the frame of tags or fusion proteins. Addgene will also align our NGS results with the full or partial sequences provided by the depositing lab.
As part of our QC processing, Addgene will contact the depositing laboratory if there are any discrepancies between their cloning data or deposited sequences and our NGS results prior to making plasmids available online. When the analysis is complete (usually 4-6 weeks from when they are received at Addgene), the sequencing results are posted online for the community. NGS full plasmid sequence data can be found under the heading “Full Sequences from Addgene” after clicking the Sequences link on the plasmid page. All full plasmid sequences generated by Addgene are titled “Addgene NGS result”. If NGS cannot assemble a full plasmid sequence, but provides useful data, Addgene will make this available as a partial sequence labelled as “Addgene Partial NGS Result.” If during validation of a plasmid you received from Addgene you identify a sequence discrepancy relative to the provided sequences, please report this problem to our Technical Support Team, and they would be happy to provide assistance to resolve the issue.