Patent SG-11201808675V-A (SG)
Title: Method and kit for the generation of dna libraries for massively parallel sequencing
Inventor: MANARESI NICOLÒ
IPC: C12Q1/68
Filed: 2017-04-14
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property -, Organization 11M111111111101111011111 0101111101110101111111011111H101111111111110111111 International Bureau 0.. .... .. ..... ..or::,„, (10) International Publication Number (43) International Publication Date WO 2017/178655 Al 19 October 2017(19.10.2017) WIPO I PCT (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every C12Q 1/68 (2006.01) kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, (21) International Application Number: BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, PCT/EP2017/059075 DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (22) International Filing Date: HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, 14 April 2017 (14.04.2017) KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, (25) Filing Language: English NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, (26) Publication Language: English RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, (30) Priority Data: ZA, ZM, ZW. 102016000039154 15 April 2016 (15.04.2016) IT (84) Designated States (unless otherwise indicated, for every (71) Applicant: MENARINI SILICON BIOSYSTEMS kind of regional protection available): ARIPO (BW, GH, S.P.A. [IT/IT]; Via Giuseppe di Vittorio, 21B/3, 40013 GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, CASTEL MAGGIORE (IT). TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, (72) Inventors: MANARESI, Nicoll); Via Santo Stefano, 132, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, 40125 BOLOGNA (IT). BUSON, Genny; c/o MENARINI DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SILICON BIOSYSTEMS S.p.A., Via Giuseppe di Vit- SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, torio, 21B/3, 40013 CASTEL MAGGIORE (IT). GW, KM, ML, MR, NE, SN, TD, TG). _ TONONI, Paola; c/o MENARINI SILICON BIOSYS- — TEMS S.p.A., Via Giuseppe di Vittorio, 21B/3, 40013 Published: — CASTEL MAGGIORE (IT). — — with international search report (Art. 21(3)) = (74) Agents: BOSIA, Alessandra et al.; STUDIO TORTA 10121 TORINO S.p.A., Via Viotti, 9, (IT). = = = (54) Title: METHOD AND KIT FOR THE GENERATION OF DNA LIBRARIES FOR MASSIVELY PARALLEL SEQUEN- CING = _ SSONN _ NSB ~6 = 566 = = VMS = ' 1 - R -2PRSBC ' PPR33 .,..- , ' - ' ' , SS / - , , Figure 3 1PR / - BC k / Il , 11 in 8SLINA 11 in ,tD GC Ir---- (57) : There is disclosed a method of generating a massively parallel sequencing library comprising the steps of :a) provid- ing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adaptor;b)re-amp - 11------ lifying the primary WGA DNA library using at least one first primer (1PR) and at least one second primer (2PR); the at least one 1-1 C first primer (1PR) comprising from 5' to 3' at least one first sequencing adaptor (1PR5SA), at least one first sequencing barcode N (1PR5BC) and a first primer 3' section (1PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse 0 complementary; the at least one second primer (2PR) comprising from 5' to 3' at least one second sequencing adaptor (2PR5SA) dif - ferent from the at least one first sequencing adaptor (1PR5SA), and a second primer 3' section (2PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary.