5500 Series Genetic Analyzers: Reagents and Consumables ORDERING INFORMATION Publication Part Number 4465650 Rev. D Revision Date April 2012 Use this guide to select the reagents and consumables required to perform a sequencing run on the 5500 Series Genetic Analyzers. • Select Sequencing Primer (SP) kit(s), as required by the library type and desired analysis tag. In the following illustrations, primer names are shown above the arrows that represent the direction of the sequencing read generated by the primers, and the analysis tag is shown in parentheses. • Select forward or reverse Sequencing Reagents (SR), as appropriate for the sequencing run type chosen. • Select other reagents and consumables required for all sequencing runs. • (Optional) Select Exact Call Chemistry (ECC) reagents, if an ECC primer round (forward only) will be included. FWD1 (F3) Fragment REV1 DNA (F5-DNA) FWD2 (BC) P1 DNA FWD1 (F3) Mate-paired P1 Mate-pair tag IA BC FWD1 (F3) P2 Whole transcriptome FWD2 (R3) IA Mate-pair tag F3 (forward tag) (Part no.) F5-DNA or F5-RNA (reverse tag) (Part no.) Small RNA or SAGE™ P2 R3 (mate-paired tag) or BC (barcode tag) (Part no.) Forward-only SOLiD® FWD1 SP Kit (4463011) — Forward-only, multiplex SOLiD® FWD1 SP Kit (4463011) — Paired-end SOLiD® FWD1 SP Kit (4463011) SOLiD® REV1 (DNA) SP Kit (4463020) Paired-end, multiplex SOLiD® FWD1 SP Kit (4463011) SOLiD® REV1 (DNA) SP Kit (4463020) SOLiD® FWD2 SP Kit (4463012) — SOLiD® FWD2 SP Kit (4463012) — • • SOLiD® FWD2 SP Kit (4463012) — • • • • DNA mate-paired libraries SOLiD® FWD1 SP Kit (4463011) • • RNA whole transcriptome libraries Forward-only SOLiD® FWD1 SP Kit (4463011) — Forward-only, multiplex SOLiD® FWD1 SP Kit (4463011) — Paired-end SOLiD® FWD1 SP Kit (4463011) SOLiD® REV1 (RNA) SP Kit (4463019) Paired-end, multiplex SOLiD® FWD1 SP Kit (4463011) SOLiD® REV1 (RNA) SP Kit (4463019) — SOLiD® Small RNA SP Kit (4463013) — Forward-only, multiplex SOLiD® Small RNA SP Kit (4463013) — • • SOLiD® FWD2 SP Kit (4463012) — SOLiD® FWD2 SP Kit (4463012) RNA small RNA or SAGE™ libraries Forward-only cDNA — SOLiD® FWD2 SP Kit (4463012) P1 • • • • IA BC P2 FWD2 (BC) cDNA Forward or reverse Sequencing Reagent (SR) kits and buffers (Part no.) DNA fragment libraries Mate-paired P1 Small RNA (F3) Sequencing Primer (SP) kits by analysis tag Sequencing run REV1 RNA (F5-RNA) FWD2 (BC) IA BC P2 Reagents and consumables for all run types (Part no.) Required • SOLiD® Pre Deposition Plus SR S75/S50 Kit (4472967) SOLiD® FWD Buffer • SOLiD® FlowChip Deposition (4459193) Buffer 1 (4462407) • SOLiD® FlowChip, 6-Lanes (4461826,1-pk) SOLiD® FWD SR S75/S50 Kit • SOLiD® Buffer D (Reset) SOLiD® FWD Buffer (4458935) (4459193) SOLiD® REV SR Kit (4463030) • SOLiD® Run Cycle Buffer Kit (4461840) SOLiD® REV Buffer (4463027) – SOLiD® Buffer A (4463024) – SOLiD® Buffer B (4463021) SOLiD® FWD SR S75/S50 Kit – SOLiD® Buffer C (4458932) SOLiD® FWD Buffer – SOLiD® Buffer E (4463016) (4459193) – SOLiD® Buffer F v2 (4476391) – SOLiD® Buffer G (4458933) ® SOLiD FWD SR S75/S50 Kit (Optional) Exact Call Chemistry SOLiD® FWD Buffer reagents, forward (4459193) SOLiD® FWD ECC Module (4463032; see the 5500 Series SOLiD® FWD SR S75/S50 Kit Genetic Analyzers User Guide) SOLiD® FWD Buffer (Optional) Workflow Analysis (4459193) reagents SOLiD® REV SR Kit (4463030) SOLiD® BA Primer, 2-pack SOLiD® REV Buffer (4463027) (4472034) or SOLiD® Sequencing QC, 2-pack (4472035) SOLiD® FWD Kit† • SOLiD® FWD SR S75/S50 Kit • SOLiD® FWD Buffer (4459193) † The SOLiD® FWD SR S75 Kit is best configured for sequencing read lengths up to 75 bp for barcoded runs and is available in two sizes: Double (4473607) and Single (4473598). The SOLiD® FWD SR S50 Kit is best configured for sequencing read lengths up to 50 bp for barcoded runs, as well as sequencing read lengths up to 60 bp with no barcodes, and is available in two sizes: Double (4473606) and Single (4473595). 5500 Series Genetic Analyzers: Reagents and Consumables Quick Reference 04/2012 1 Kit unit definitions Category FlowChip and deposition Sequencing primer kits Kit Category 1 kit SOLiD® FlowChip Deposition Buffer 1 (4462407) 6 lanes SOLiD® FlowChip, 6-Lanes (4461826) SOLiD® Kit Unit SOLiD® FWD SR S75 Kit, Double (4473607) 750–1032 cycles† SOLiD® FWD SR S75 Kit, Single (4473598) 375–516 cycles 1 each SOLiD® FWD SR S50 Kit, Double (4473606) 500–732 cycles 12 lanes SOLiD® FWD SR S50 Kit, Single (4473595) 250–366 cycles Small RNA SP Kit (4463013) 12 lanes SOLiD® 900–1212 cycles† SOLiD® REV1 (DNA) SP Kit (4463020) 12 lanes SOLiD® FWD ECC Module, Double (4463032) 120–180 cycles† SOLiD® REV1 (RNA) SP Kit (4463019) 12 lanes Reverse probes/ enzymes, buffer SOLiD® REV SR Kit, Double (4463030) 280–420 cycles† SOLiD® 240–420 cycles† Buffers for sequencing in all directions SOLiD® Run Cycle Buffer Kit, Double (4461840) 950–1632 cycles† SOLiD® Buffer D, Double (Reset) (4458935) 216 primer resets† SOLiD® SOLiD® Workflow analysis kits Unit SOLiD® Pre Deposition Kit (4472967) FWD1 SP Kit (4463011) FWD2 SP Kit (4463012) 12 lanes SOLiD® BA Primer, 2-pack 24 lanes SOLiD® Sequencing QC, 2-pack 24 lanes Forward probes/ enzymes, buffer FWD Buffer, Double (4459193) REV Buffer, Double (4463027) † Double sizes support 12 simultaneous lanes of 75 nt forward + 35 nt reverse + 10 nt barcode (same library type). If a full FlowChip is not run, fewer lanes or cycles are supported, due to dead volumes required by the sequencer. 5500 Series Genetic Analyzers reagent placement Kit SOLiD® SOLiD® Component FWD1 SP Kit SOLiD® FWD1 Seq. Primers FWD2 SP Kit SOLiD® FWD2 Seq. Primers SOLiD® Small RNA SP Kit SOLiD® Small RNA Seq. Primers SOLiD® REV1 (DNA) SP Kit SOLiD® REV1 (DNA) Seq. Primers SOLiD® REV1 (RNA) SP Kit SOLiD® REV1 (RNA) Seq. Primers Any SP kit SOLiD® Mixing Tubes Any SP kit, or SOLiD® Sequencing QC 2-pack SOLiD® Sequencing QC SOLiD® FWD1 SP Kit SOLiD® Small RNA SP Kit SOLiD® REV1 (DNA) SP Kit SOLiD® REV1 (RNA) SP Kit SOLiD® BA Primer, 2-pack SOLiD® BA Primer† SOLiD® FWD ECC Module SOLiD® FWD ECC Seq. Primers† Location Reagent deck Reagent strip groups • FWD SR: Forward sequencing and forward ECC reagents • MT: Mixing tubes • REV SR: Reverse sequencing reagents • SP: Sequencing primers • SQC: Sequencing QC A RV SOLiD® FWD ECC Seq. Reagents Kit SOLiD® SOLiD® Location Kit FWD SR S75/S50 Kit SOLiD® Buffer A FWD Buffer SOLiD® Buffer B SOLiD® REV SR Kit 11a 11b SOLiD® REV Buffer C B E FD D SP MT REV SR Location 7 8 9 10a 10b SQC 11a 11b FWD SR SOLiD® Buffer C SOLiD® Buffer E — SOLiD® Buffer D (Reset) F SOLiD® Buffer F v2 G 1 2 3 4 5 6a 6b SOLiD® Buffer G † Use the SOLiD® BA Primer component for sequencing runs that do not include Exact Call Chemistry (ECC). For sequencing runs that include ECC, do not use the SOLiD® BA Primer component. Instead, use the SOLiD® FWD ECC Seq. Primers strip from the SOLiD® FWD ECC Module. (The SOLiD® FWD ECC Seq. Primers include the SOLiD® BA Primer.) For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. LIFE TECHNOLOGIES CORPORATION AND/OR ITS AFFILIATE(S) DISCLAIM ALL WARRANTIES WITH RESPECT TO THIS DOCUMENT, EXPRESSED OR IMPLIED, INCLUDING BUT NOT LIMITED TO THOSE OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE, OR NON-INFRINGEMENT. TO THE EXTENT ALLOWED BY LAW, IN NO EVENT SHALL LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) BE LIABLE, WHETHER IN CONTRACT, TORT, WARRANTY, OR UNDER ANY STATUTE OR ON ANY OTHER BASIS FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING BUT NOT LIMITED TO THE USE THEREOF. NOTICE TO PURCHASER: PLEASE REFER TO THE 5500 Series Genetic Analysis Systems Limited Use Label Licenses (PUBLICATION PART NO. 4465100) AND THE 5500 Series Genetic Analyzers User Guide (PUBLICATION PART NO. 4456991) FOR LIMITED LABEL LICENSE OR DISCLAIMER INFORMATION. © 2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. 2 5500 Series Genetic Analyzers: Reagents and Consumables Quick Reference 04/2012