Sequencing of tardigrade cDNA amplicons from bacterial expression plasmids
To identify tardigrade proteins whose expression could improve bacterial (E. coli) desiccation survival, we cloned tardigrade cDNAs into a bacterial expression plasmid (pDest17). After inducing protein expression with 0.2% L-arabinose, bacteria were either desiccated or kept as non-desiccated controls. The cDNA inserts of the plasmids were isolated and amplified by low-cycle (12x) PCR with a Kapa2G Robust Hotstart PCR kit (Roche, Ref #07961073001). PCR amplicons were cleaned with a ChargeSwitch-Pro PCR cleanup kit (Invitrogen, Cat #CS32050) and sonicated to ~200 bp. Sequencing libraries were prepared with a Kapa DNA hyper kit. Batches were pooled onto lanes for paired end 75 bp read sequencing on an Illumina HiSeq4000.