I don’t have access to a Covaris instrument. Can I use alternative fragmentation methods?
We have evaluated only Covaris fragmented DNA during the development of the Ovation Ultralow System V2. Other sonication instruments or enzymatic fragmentation may be suitable but will need to be optimized.
Does Tecan provide reagents for performing the fragmentation step of the protocol?
Tecan does not provide the reagents used in the fragmentation steps. We suggest the Covaris instrument be utilized for DNA fragmentation, as suggested in the “materials” section of the User Guide.
Can I modify the number of PCR amplification cycles recommended by the Ovation Ultralow System V2 workflow when using different DNA input amounts?
Generally speaking, fewer PCR cycles will be needed when working with larger input amounts. See Table 6 of the User Guide for guidelines on the number of cycles to use.
Can I combine the barcoded libraries prior to the PCR amplification step?
This is not recommended. The stoichiometry of barcoded libraries may be adversely affected by this modification to the workflow. We suggest that the libraries be amplified and quantified independently before being pooled for use on the sequencer.