Inactive Total Small Business Set-Aside (FAR 19.5)

AMENDMENT 6/25/2021 QUESTION: May I assume that you will be sending gDNA as starting material? ANSWER: The submitted material will be DNA. AMENDMENT 6/25/2021 QUESTION: We have used both primer sets i... AMENDMENT 6/25/2021 QUESTION: May I assume that you will be sending gDNA as starting material? ANSWER: The submitted material will be DNA. AMENDMENT 6/25/2021 QUESTION: We have used both primer sets in our lab frequently and can perform all of the necessary steps to generate sequencing data as well as meet requested timetables. The RFQ does not state what sequencing depth is required, but we should be able to run all amplicons on a single MiSeq 2x300 run, which would provide about 20k reads per sample. If greater sequencing depth is required or additional PCR replicates, please inform. ANSWER: 20K reads per sample are fine. The SOW/SON indicated the total # of samples but did not specify the # of samples per submission as we don't know that yet. If this quote accurate for more than a single submission? For example, we know that the 16S samples will not be submitted with the CO1 samples. Often there is a pricing scale depending on the # of samples submitted. AMENDMENT 6/24/2021 QUESTIONS: "To be summarized, you want to do 16s rRNA sequencing for 200 DNA samples and COI sequencing for 60 samples, right? Please confirm. We will use one MiSeq lane for these 260 samples. For COI sequencing, Our supplier is not sure why you want to do two PCR rounds, actually, one PCR using BF2+BR2 with in-line tags & Illumina sequencing adaptors will be good enough, as we will do with the 16s sequencing, please let me know if this is fine with you." ANSWER: Nope, that isn't fine for the CO1. We are following the protocol of the reference cited in the tech specs, which uses a 2-PCR process to reduce tag switching that occurs with the single PCR. First step has no tags, while the 2nd step does. This is recommended by colleagues working with similar taxa. COMBINED SYNOPSIS/SOLICITATION PURCHASE OF 200 16S RRNA MICROBIAL AND 60 CYTOCHROME C OXIDASE 1 SAMPLES HIGH THROUGHPUT SEQUENCING FOR LOWER TROPHIC LEVELS IN MARINE WATERS IN ACCORDANCE WITH THE ATTACHED STATEMENT OF NEED (I) This is a combined synopsis/solicitation for commercial items prepared in accordance with the format in FAR Subpart 12.6, as supplemented with additional information included in this notice and in accordance with the simplified acquisition procedures authorized in FAR Part 13. This announcement constitutes the only solicitation; quotes are being requested and a written solicitation will not be issued. (II) This solicitation is issued as a request for quotation (RFQ). Submit written quotes on RFQ Number NFFP72102102687SRG. (III) The solicitation document and incorporated provisions and clauses are those in effect through Federal Acquisition Circular 2021-05 (MAR 2021) (DEVIATION 2020-11) (AUG 2017). (IV) This solicitation is being issued as a Total Small Business Set-Aside. The associated NAICS code is 334516. The small business size standard is 1000 employees. (V) This combined solicitation/synopsis is for purchase of the following commercial services: CLIN 0001: Purchase of a quantity of 200 samples, 16S rRNA microbial, Primer of 515fB - GTGYCAGCMGCCGCGGTAA; 806rB – GGACTACNVGGGTWTCTAAT (updated EMP pair), Amplicon size of 292 bp and reference is Earth Microbiome Project 16S Illumins Protocol (https://earthmicrobiome.org/protocols-and-standards/16s/) $__________ per sample and extended price for 200 samples $__________. CLIN 0002: Purchase a quantity of 60, Cytochrome c oxidase 1, primer of BF2 - GCHCCHGAYATRGCHTTYCC BR2 - TCDGGRTGNCCRAARAAYCA, amplicon size of 424 bp and reference of Elbrecht & Steinke, 2019. Note this is a 2-PCR library prep with 1st PCR for amplification using unmodified BF2 + BR2, & 2nd PCR using BF2+BR2 with in-line tags & Illumina sequencing adaptors. No cleanup between 1st & 2nd PCR. Will need to synthesize ten (10) primers: unmodified BF2, unmodified BR2, four (4) modified BF2, four (4) modified BR2 (Fig 3 of reference) $__________ per sample and extended price for 60 samples $__________. Total of both CLIN 0001 & 0002 $___________________