Q and A.docx

ABL-22067

Q&A

The key tasks explicitly asks for 8 lanes of NovaSeq S4 sequencing (i.e., two S4 flow cells) with a yield of 800 Gb of sequence data per lane. Using the standard expected Gb output provided by Illumina, you may only get 2400-3000 Gb per flow cell or about 600-750 Gb per lane. For the quotation, would you like to stick to 8 lanes, which should yield ~600-750 Gb per lane, or increase the number of lanes/flow cells being used to try to ensure 800 Gb per lane target (or some other target based on the number of libraries)? By our estimates, you could go up to 3 or 4 full S4 flow cells and stay well under the $150,000 simplified acquisition threshold.

We would like to keep the same number of lanes and are ok with 600-750gb yield per lane.

Please clarify on the second task regarding QC library preparation. We are interpreting this to mean that you will send 16 already prepared libraries and would like a QC analysis on the libraries we receive, including, for example, a fragment analysis and DNA quantitation (e.g. QuBit) to ensure libraries are high quality before sequencing. Is this accurate?

Yes

The description and deliverable sections reference Illumina NovaSeq 6000 sequencing, using PE300 (paired end, 300bp) chemistry. I think this may be a typo as the S4 flow cells are only capable of PE150 run mode. The combined data from either side of the library would of course be 300 bp but not in a paired-end configuration as stated in the solicitation.

Sorry yes we mean PE150 chemistry 300 cycle kits

Is it okay to assume that the lab is looking for 300 bp data from PE150 S4 flow cells?

Yes