U. S. Environmental Protection Agency, Contracts Management Division (MD-33), Attn: ORDSC, Research Triangle Park, NC 27711

A -- MOLECULAR AND ANALYTICAL SERVICES-GULF ECOLOGY DIVISION SOL Q81001 DUE 120298 POC Sya Mayes (919) 541 3416 WEB: click here for web page, http://www.epa.gov/oam.rtp_cmd. E-MAIL: click here to contact the contract specialist via, mayes.sya@epa.gov. THIS IS A COMBINED SYNOPSIS/SOLICITATION FOR COMMERCIAL ITEMS PREPARED IN ACCORDANCE WITH THE FORMAT IN SUBPART 12.6, AS SUPPLEMENTED WITH ADDITIONAL INFORMATION INCLUDED IN THIS NOTICE. THIS ANNOUNCEMENT CONSTITUTES THE ONLY SOLICITATION; QUOTES ARE BEING REQUESTED AND A WRITTEN SOLICITATION WILL NOT BE ISSUED. The solicitation number is Q81001 and the solicitation is being issued as a Request for Quote (RFQ). The solicitation document and incorporated provisions and clauses are those in effect through Federal Acquisition Circular 97-05. The Environmental Protection Agency (EPA) anticipates that the contract will be awarded as a result of full and open competition. A firm-fixed-price contract is anticipated to result from the award of this solicitation. The associated Standard Industrial Code (SIC) is 8734. This procurement is for Differential Display Analysis of RNA isolated from the livers of fish exposed to three chemicals known to impair growth. After reamplification by the contractor the EPA willclone the samples into DNA which will be returned to the contractor for sequencing. Oligonucleotides of high purity shall be synthesized according to EPA specifications and be used as primers for bacterial DNA sequencing and PCR amplification of various DNA from the environment and other sources (e.g. plasmids). High purity monoclonal antibodies, to be used as specific serum or cellular biomarkers, will be raised against antigens (fish or bacterial cell wall proteins) provided by the EPA. The contractor must demonstrate their expertise in each of the proposed areas. The contractor is required to include a copy of their Quality Assurance/Quality Control protocols with their submission. The contractor must permit a QA audit of laboratory and/or data entry procedures by an authorized agent of EPA at any time during the contract period (given advanced notification). Results of the Differential Display Analysis and DNA Sequencing will be provided in both electronic and hard copy to the EPA Project Officer. Specific antibodies and oligonucleotides shall be delivered to the EPA Project Officer by an overnight delivery service upon completion. A written report detailing the methods used and QA measures applied shall also be provided by the contractor. Original records such as laboratory notebooks, reproductions/photographs of Western blot gels or ELISA plates, and X-ray film shall be retained by the contractor for a period of three years following the submission of the final report. THE CONTRACTOR SHALL PROVIDE EPA WITH A COST ESTIMATE ON A PER SAMPLE BASIS FOR EACH OF THE FOLLOWING LINE ITEMS. Item 01: Differential Display RT-PCR (DD-RT-PCR). The EPA will prepare the RNAs, proteinase K and DNase treat the RNA, check the RNA by spectrophotometer at 260 and 280 nm to determine the quantity and an agarose check gel to determine the the quality of the RNA. The contractor shall confirm the quality of the RNA by agarose gel and perform DD-RT-PCR on the samples using the primer pairs suggested by the EPA. In addition, one lane to determine the presence of DNA shall accompany each assay. The EPA shall be consulted to select the differentially expressed bands from the DD gels and the service laboratory shall reamplify each band and check the reamplification by agarose gel electrophoresis. The contractor shall provide EPA with the reamplified bands for cloning and for verification of differential expression by Northern blot analysis. The EPA will then provide the service laboratory with 5 clones from each amplified band for DNA sequence analysis. Number of DD gels: 30. Number of primer pairs: 20. Number of reamplified bands: 300. Item 02: DNA Sequencing. The service laboratory shall perform automated DNA sequencing using single or double stranded DNA (plasmid or PCR product) templates supplied by the EPA. Reactions using small amounts of template to be accomplished by thermocycler. The purity and concentration of the supplied template is to be checked spectrophotmetrically and by agarose gel electrophoresis by the EPA. EPA will provide a photo of the agarose gel. The service laboratory shall perform dideoxynucleotide terminated sequencing reactions with fluorescent dye-labeled primers or terminators according to the sequencing kit manufacturer's protocol. For reactions extended from unique priming sites, labeled primers, unlabeled primers or primer sequences shall be provided by the requestor. The service laboratory shall provide ultracold (-70o C) long term storage of supplied templates and primers. Sequencing of large pieces of DNA by primer walking shall be available as an option. The EPA will indicate which samples should be read in the standard 300-450 bases of interpretable sequence and which samples should be handled for 450-600 bases of interpretable sequence. Data shall be provided on diskette and hard copies as requested. Turn around time not to exceed 30 days. Number of Samples: 2000. Line Item 03: Oligonucleotide Synthesis. The contractor shall provide high purity synthetic oligonucleotides. Synthesis shall allow for mixed nucleotide positions, modified bases, amino linkers and other state-of-the-art syntheses. The contractor shall maintain a demonstrably high level of quality control. Oligonucleotides shall be provided dried, prepared for dilution, and suited for use in most applications without further purification. Information provided by the contractor for each nucleotide shall include at least the amount and sequence of the dried product. Turn around time should be less than five days, with next day service as an option. Quantity: 500 oligonucleotides of about 20 nucleotides in length. Line Item 04: Monoclonal Antibody Production. The EPA will provide 1 to 3 mg of purified antigen for each antibody project. The contractor shall immunize 4 different mice at 3 to 4 time intervals, collect blood from the tail vein to check for antibody production to the antigen by ELISA (preferably) or Western blot, conduct up to 2 fusions, plate out the resulting hybridomas and screen the fusion supernatants by ELISA (preferably) or Western blot. Provide for 2 rounds of screening if necessary. Five wells shall be selected and cloned, followed by another round screening. The contractor shall provide 50 to 100 ml of supernatant from each clone, isotype the clones and check for mycoplasma. One of the clones shall be selected for mass production of antibody to generate 50 mg of purified antibody (purified by Protein G). Quantity: 6 Antibody projects. Offers will be evaluated using the following technical capabilities criteria: 1.) QA/QC protocols; 2.) Demonstrated analytical experience with proposed analytical techniques; 3.) Demonstrated previous experience with fish tissue samples; 4.) Proposed methodology; 5.) Experience with similar size/scope projects; 6.) Past performance -- provide 3 references; 7.) Report preparation and records management; and 8.) Adhering to deadlines. Award will be made to the lowest priced offer meeting all of the technical requirements. The following FAR provisions apply to this solicitation: 52.212-1, Evaluation-Instructions to Offerors-Commercial; 52.212-2, Evaluation-Commercial Item. Evaluation criteria to be included in paragraph (a) of provision 52.212-2 are as follows: (i) technical capability to meet the Government requirements; (ii) past performance; (iii) price, in decreasing order of importance. Technical and past performance, when combined, are significantly more important than price. All offerors are to include with their offers a completed copy of provision 52.212-3, Offeror Representations and Certifications-Commercial Items. The following FAR clause applies to this acquisition: 52.212-4, Contract Terms and Conditions-Commercial Items. The following additional FAR clauses which are cited in clauses 52.212-5 are applicable to the acquisition: 52.203-6, Restrictions on Subcontractor Sales to the Government, with Alternate I; 52.203-10, Price or Fee Adjustment for Illegal or Improper Activity; 52.219-14, Limitation on Subcontracting; 52.222-26, Equal Opportunity, 52.222-35, Affirmative Action for Special Disabled and Vietnam Era Veterans; 52.222-36 Affirmative Action for Handicapped Workers; 52.222-37, Employment Reports on Special Disabled Veterans and Veterans of the Vietnam Era; 52.225-3, Buy American Act -- Supplies; 52.222-41, Service Contract Act of 1965, As Amended. Cost and technical information should be submitted in separate proposals. Please submit two copies of technical propopals and one copy of cost proposal to Sya M. Mayes, Contract Specialist, U.S. Environmental Protection Agency, MD-33, Research Triangle Park, NC 27711. Courier Delivery Address is: U.S. Environmental Protection Agency, Receptionist-Administration Bldg. Lobby, Attn: Sya M. Mayes, 79 Alexander Drive, Research Triangle Park, NC 27709. All offers are due by December 2, 1998, 4:00 p.m. ET. Offers/bids received after the exact time specified will not be considered. No telephonic or faxed requests will be honored. Posted 11/17/98 (W-SN272261). (0321)

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