CÔNG TY CP CÔNG NGHỆ SINH HỌC BIONET VIỆT NAM Số 12N1 - Ngõ 53 – Đường Hoàng Cầu – Phường Ô Chợ Dừa – Quận Đống Đa – Hà Nội Tel: 04-66-747-102; Hotline: 0988 683 082; Email:support@bionet.vn BÁO CÁO CÔNG VIỆC THEO TUẦN Người thực hiện: Vũ Anh Tuấn Tuần 22 – 29/9/2014 I Các công việc làm tuần STT Công việc Ghi Đọc tài liệu Đột biến G6PD Thiết kế Primer cho đột biến thường gặp Việt Nam Kết cụ thể liệt kê Viangchan 871G/A Phenotype: G6PD VIANGCHAN (Khác: G6PD JAMMU) cDNA Nucleotide : 871 GA Exon: Codon: Val291Met Variation ID: rs137852327 RefSeqGene Mapping: NG009015.2 Gene (ID): G6PD (2539) Position: 19451 F_inner(G): 5’-GTCCGTGATGAGAAGGTCACGG-3’ R_inner(A): 5’-TGCACCTCTGAGATGCATTTCAAAAT-3’ F_common: 5’-CCAGATCTACCGCATCGACCACTA-3’ R_common: 5’-ATCATGGAACTGCAGCCTCACCT-3’ 22bp 22bp 24bp 24bp GC% GC% GC% GC% 59.1 38.5 54.2 52.5 Tm:63.5 Tm:62.6 Tm:64.2 Tm:64.7 M – Product: 317 bp N –Product: 277 bp Common: 544 bp Kaiping 1388G/A Phenotype: G6PD KAIPING (Khác: G6PD ANANT, G6PD DHON, G6PD PETRICK-LIKE, G6PD SAPPORO-LIKE) cDNA Nucleotide : 1388 GA Codon: Arg463His Exon: 12 Variation ID: rs72554664 RefSeqGene Mapping: NG009015.2 Gene (ID): 2539 F_inner(A): 5’-GAGCTCCGTGAGGCCTGTCA-3’ R_inner(G): 5’-TGCAGCAGTGGGGTGAAAAGAC-3’ F_common: 5’-GAGTCGGAGCTGGACCTGACCTAC-3’ R_common: 5’-TCTCTTCATCAGCTCGTCTGCCTC-3’ M – Product: 127 bp N –Product: 161 bp Common: 246 bp 20bp 22bp 24bp 24bp Position: 20316 GC% GC% GC% GC% 65.0 54.5 54.2 62.5 Tm:64.6 Tm:64.1 Tm:66.4 Tm:64.6 3.Canton1376 G/T cDNA Nucleotide: 1376 GT Codon: Arg459Leu Exon: 12 Variation ID: rs72554665 Position: 20304 F_inner(G): 5’-CGCAGCGACGAGCTTCG-3’ R_inner(T): 5’-GGTGAAAATACGCCAGGCCTTAA-3’ F_common: 5’-GGAGCTGGACCTGACCTACGG-3’ R_common: 5’-TTCATCAGCTCGTCTGCCTCC-3’ 17 23 21 21 70.6 47.8 66.7 57.1 62.2 62.0 64.5 62.8 M-Product: 145 bp N-Product: 132 bp Common: 237 bp 4.Union1360 C/T cDNA Nucleotide: 1360 CT gDNA : 13763 Codon: Arg454Cys Exon: 11 Class : II Variation ID: rs137852324 Position: 20184 (Sử dụng primer Nga báo cáo) Normal primer F-5’-TTCTGCGGGAGCCAGATGCACTTCGTGC-3’ ACG Mutant primer F-5’-TTCTGCGGGAGCCAGATGCACTTCGTTT-3’ Reverse primer (common) R-5’-AGTGACGGGTGGAGGAGAGGCATGAGGTA-3’ Reverse complement 5’-TACCTCATGCCTCTCCTCCACCCGTCACT-3’ Mismatch: TG (weak), CT (strong) Kiểm tra NCBI/Primer-BLAST :glucose-6-phosphate 1-dehydrogenase Sequence (5'->3') Length Start Stop Tm GC% Forward primer TTCTGCGGGAGCCAGATGCACTTCGTGC 28 15170 15197 72.53 60.71 Reverse primer AGTGACGGGTGGAGGAGAGGCATGAGGTA 29 15490 15462 71.22 58.62 Product length 321 PCR Product: TTCTGCGGGAGCCAGATGCACTTCGTGCGCAGGTGAGGCCCAGCTGCCGGCCCCTGCATACCTGTG GGCTATGGGGTGGCCTTTGCCCTCCCTCCCTGTGTGCCACCGGCCTCCCAAGCCATACCATGTCCCCT CAGCGACGAGCTCCGTGAGGCCTGGCGTATTTTCACCCCACTGCTGCACCAGATTGAGCTGGAGAA GCCCAAGCCCATCCCCTATATTTATGGCAGGTGAGGAAAGGGTGGGGGCTGGGGACAGAGCCCAGC GGGCAGGGGCGGGGTGAGGGTGGAGCTACCTCATGCCTCTCCTCCACCCGTCACT Mediterranean 563 C/T cDNA Nucleotide : 563 GC Codon: Ser188Phe Exon: Variation ID: rs5030868 RefSeqGene Mapping: NG009015.2 Gene (ID): 2539 F_inner(G): 5’-ACCGGCTGTCCAACCACATATC-3’ R_inner(C): 5’-CCTCACGGAACAGGGCGA-3’ F_common: 5’-CACATGAATGCCCTCCACCTG-3’ R_common: 5’-CCAAAGGGCTCCTTGAAGGTG-3’ Position: 18154 22 18 21 21 54.5 66.7 57.1 57.1 63.2 62.3 61.9 61.8 22 19 22 22 54.5 57.9 54.5 54.5 62.7 69.3 62.8 62.9 20 21 25 26 65.0 57.1 48.0 38.5 62.0 62.4 63.6 59.6 21 19 23 23 52.4 63.2 47.8 56.5 61.2 62.4 61.7 63.0 M-Product: 185 bp N-Product: 211 bp Common: 356 bp Mahidol 487 G/A cDNA Nucleotide: 487 GA Codon: Gly163Ser Exon: Variation ID: rs137852314Position: 18078 F_inner(G): 5’- GAGTCCTGCATGAGCCAGAGAA-3’ R_inner(A): 5’- CGATGATGCGGTTCCATCC-3’ F_common: 5’- GGTGACCTGGCCAAGAAGAAGA-3’ R_common: 5’- TAGGTGGTTCTGCATCACGTCC-3’ M-Product: 328 bp N-Product: 385 bp Common: 672 bp 7.Coimbra 592C/T cDNA Nucleotide: 592 CT gDNA : 11763 Codon: Arg198Leu Exon: Class II Variation ID: rs137852330 Position: 18183 F_inner(C): 5’-CCGTGAGGACCAGATCTCCC-3’ R_inner(T): 5’-GCCCAGGTAGTGGTCGATTCA-3’ F_common: 5’-GTCACCAAGAACATTCACGAGTCCT-3’ R_common: 5’-GATGATCCCAAATTCATCGAAATAGC-3’ M-Product: 197 bp N-Product: 165 bp Common: 321 bp 8.Vanua Lava 592C/T cDNA Nucleotide: 592 CT gDNA : 10884 Codon: Leu127Pro Exon: Class II Variation ID: rs1050829 (????) Position: 17303 F_inner(C): 5’-CAACAGCCACATGAATGCACC-3’ R_inner(T): 5’-GCCTGTGACCCCAGGTTGA-3’ F_common: 5’-TGGCCAAGAAGAAGATCTACCCC-3’ R_common: 5’-GGTAGTGGTCGATGCGGTAGATC-3’ M-Product: 274 bp N-Product: 246 bp Common: 480 bp BaoLoc 352 T/C cDNA Nucleotide: 352 TC gDNA: 10853 Codon: Tyr118His Exon: Class II Variation ID: rs1050829(?????) Position: F_inner(T): 5’-GCCAGTACGATGATGCAGCCTACT-3’ R_inner(C): 5’-GGCTGTTGAGGCGCTGTTG-3’ F_common: 5’-GGGTGACCTGGCCAAGAAGAA-3’ R_common: 5’-AGGTAGTGGTCGATGCGGTAGATCT-3’ 24 19 21 25 54.2 63.2 57.1 52.0 65.2 62.8 62.9 65.3 M-Product: 251 bp N-Product: 281 bp Common: 489 bp Primer kiểm tra tính đặc hiệu NCBI/Blast II Kế hoạch làm việc tuần (những công việc giao kế hoạch cá nhân đề ra) STT Công việc Chỉnh sửa lại primer thiết kế Thiết kế cho Multiplex PCR theo thứ tự: ViangChan; Kaiping; Canton; Union; Mediterranean; Coimbra; Mahidol; Vanua Lava; BaoLoc Ghi III Một số kiến nghị, góp ý, sáng kiến, đề xuất trình ban lãnh đạo xem xét ………………………………………………………………………………………………… ……………………………………………………………………… IV Nhận xét, góp ý đạo TGĐ ………………………………………………………………………………………………… ………………………………………………………………………………………………… ………………………………………………………………………………………………… Hà Nội, Ngày 29 tháng năm 2014 Người báo cáo Vũ Anh Tuấn ... Position: 181 54 22 18 21 21 54. 5 66.7 57.1 57.1 63.2 62.3 61.9 61.8 22 19 22 22 54. 5 57.9 54. 5 54. 5 62.7 69.3 62.8 62.9 20 21 25 26 65.0 57.1 48 .0 38.5 62.0 62 .4 63.6 59.6 21 19 23 23 52 .4 63.2 47 .8 56.5... 5’-TTCATCAGCTCGTCTGCCTCC-3’ 17 23 21 21 70.6 47 .8 66.7 57.1 62.2 62.0 64. 5 62.8 M-Product: 145 bp N-Product: 132 bp Common: 237 bp 4. Union1360 C/T cDNA Nucleotide: 1360 CT gDNA : 13763 Codon: Arg454Cys Exon: 11 Class... 63.2 47 .8 56.5 61.2 62 .4 61.7 63.0 M-Product: 185 bp N-Product: 211 bp Common: 356 bp Mahidol 48 7 G/A cDNA Nucleotide: 48 7 GA Codon: Gly163Ser Exon: Variation ID: rs137852314Position: 18078 F_inner(G):