multiplex pcr amplification of ancient dna

... 2008, 5:77 Figure Sensitivity of the one-step RT -PCR assay Sensitivity of the one-step RT -PCR assay Example of amplification results of ten-fold serial dilutions of in-vitro transcribed RNA H10N7 ... percentage of samples with identical matches at all five 3' terminal bases was calculated for each NA subtype Figure NPA samples One-step RT -PCR amplification of NA gene from clinical One-step RT -PCR amplification ... purified one-step RT -PCR using M13 tagged primers (e) Results of Blastn analysis (f) These samples were assayed by two-step RT -PCR before transferring to one-step RT -PCR tion of 600 μL of RLT buffer...

Ngày tải lên: 20/06/2014, 01:20

... 1000) 80 Length of local alignment and fragment length Figure Number of aligned ancient DNA fragments and average sequence length Properties of Mega BLAST alignments of ancient DNA sequences from ... accuracy of the observed number of pairwise nucleotide differences in each of these comparisons ([31] Using a model of ancient DNA fragmentation and deamination [19], we also simulated datasets of ... for informative ancient DNA shotgun sequencing Results To assess the biases introduced in the analyses of ancient DNA, we use a subset of the sequence data generated as part of the Neandertal...

Ngày tải lên: 09/08/2014, 20:22

... OPV/PPV nested multiplex PCR and confirm its applicability to viral identification and monitoring Methods and Results Multiplex PCR setting and sensitivity tests The OPV/PPV multiplex PCR was designed ... DNA manipulation, and are designed to detect specifically OPV or PPV In the present work, we report the development of a nested -multiplex PCR system for the sensitive and reliable detection of ... simulation of different combinations of several published primer pairs, using software available online [19] Two exclusive and highly conserved genes were targeted by nested -multiplex PCR: the...

Ngày tải lên: 12/08/2014, 04:20

... However, PCR using this 5'-end primer will not result in the amplification of the original coaD gene The use of the Nco I restriction site dictates what the first nucleotide of the next triplet ... triplet begins with a C and PCR amplification will change this codon from CAA to GAA (and the second residue in the recombinant protein will be glutamate instead of glutamine) An alternative ... instead of glutamine) An alternative strategy which will result in the amplification of the original gene is described in Utilisation of the Nco I site 3'-end primer No C-terminal tag is being used,...

Ngày tải lên: 14/07/2015, 23:53

... addition to rapid quantification of template RNA, real-time PCR and RTPCR offers significant advantages over standard PCR and RT -PCR for detection of DNA or RNA in terms of reduced sample handling, ... RNA Effect of incubation temperature and time on DNA degradation and RT -PCR amplification of RNA The effect of temperature on DNA degradation and RNA amplification in the presence of UNG was ... degrade carry-over amplicon DNA contamination in a sample Results Effect of UNG concentration on DNA degradation and RTPCR amplification of RNA To assess the effect of UNG on DNA degradation and RNA...

Ngày tải lên: 19/06/2014, 08:20

... addition to rapid quantification of template RNA, real-time PCR and RTPCR offers significant advantages over standard PCR and RT -PCR for detection of DNA or RNA in terms of reduced sample handling, ... RNA Effect of incubation temperature and time on DNA degradation and RT -PCR amplification of RNA The effect of temperature on DNA degradation and RNA amplification in the presence of UNG was ... degrade carry-over amplicon DNA contamination in a sample Results Effect of UNG concentration on DNA degradation and RTPCR amplification of RNA To assess the effect of UNG on DNA degradation and RNA...

Ngày tải lên: 20/06/2014, 04:20

... mixture Amplification reactions were performed in a total volume of 100 µl containing mM MgCl2, 0.2 mM each dNTPs, 20 pmol of each PCR primer, U of Taq DNA polymerase (Takara, Japan), and 10 µl of ... (7/1,682) 0.59 (10/1,682) 0.65 (11/1,682) a No of positive/No of samples examined No of negative/No of samples examined b Table Antibiotic resistance profiles of isolated E coli O157:H7 Antimicrobial ... with 0.5 µl of ethidium bromide (EtBr) per ml, visualized and photographed under UV illumination Another PCR amplification analysis was executed for confirmation of the presence of the flagellar...

Ngày tải lên: 07/08/2014, 18:21

... cloning of nonspecific or partial 16S rDNA products In general, poorer DNA profiles of bacterial species were obtained from tissue samples that gave weak PCR signals (Table 2) Bacterial 16S rDNA ... the role that this variety of intraarticular bacterial DNA plays in the pathogenesis of ReA and other forms of arthritides However, detection of bacterial DNAs in the ST of patients with arthritis ... using PCR assays with universal 16S rDNA primers, identified lower proportions of human synovial samples containing bacterial DNA: 42% of synovial fluid and ST samples in one study [18], and 10% of...

Ngày tải lên: 09/08/2014, 10:23

... purposes) PCR positivity by the broad-range PCR amplification system PCR was positive in 20 of the 27 (74.10%) patients indicating the presence of bacterial 16S rDNA within the synovial samples Of note, ... such as DNA of Pseudomonas poae, Delftia acidovorans, and Burkholderia cepacia A detailed sequence analysis of the PCR- positive samples of ReA and UA patients revealed a number of DNA of bacteria ... broad-range PCR amplification of 16S rRNA genes, cloning and sequencing of bacterial DNA For 10 minutes, 500 μl of SF were centrifuged at 10,000 g The whole SF cell pellet was subjected to DNA extraction...

Ngày tải lên: 09/08/2014, 14:22

... tube The primers and conditions used for the multiplex PCR were Page of the same as the PCR described above After multiplex PCR amplification, the PCR products were purified to remove the remaining ... each with the corresponding pair of primers, were used to amplify the promoter region and exons 18-21 of the EGFR genes (Table 1) PCR amplification of 0.2 μg DNA was performed with a denaturing ... instructions Multiplex PCR and primer extension analysis of mutations in EGFR-216 promoter region and exons 18, 19, 20, and 21 Multiplex PCR was used to amplify the promoter region and exons 18-21 of the...

Ngày tải lên: 10/08/2014, 05:21

... beacon instead of hybridization probes, 2) extraction of HBVDNA from a larger volume of serum/plasma (0.5 versus 0.2 mL), 3) use of 30 μl instead of 10 μl extracted DNA, 4) modified RTQ -PCR conditions, ... respectively, Taq DNA polymerase, dNTP mix (with dUTP instead of dTTP), and U of uracil -DNA glycosylase (Roche Applied Science Germany) The amplification profile was as follows: One cycle of initial ... Page of Analytical sensitivity of ultra sensitive RTQ -PCR To determine the analytical sensitivity of the ultra sensitive RTQ -PCR, dilutions of the WHO 1st International Standard for HBV -DNA were...

Ngày tải lên: 12/08/2014, 04:20

... question of how detection limits of chlamydial DNA might change after storage of DNA depending on the different DNA extraction methods applied To our knowledge, some laboratories performing PCR analysis ... However, the DNA extracted by either Table PCR sensitivities of the different DNA extraction methods detection Chlamydia trachomatis EB and C trachomatis PBMO DNA/ ml SF using PCR- system for amplification ... organizational aspects of the study, collection of clinical samples, culture of C trachomatis and performed DNA extraction, PCR analysis and drafted the manuscript IB and SM performed parts of DNA extraction...

Ngày tải lên: 12/08/2014, 11:22

... are suggestive of DNA damage as a cause of NS1-induced apoptosis ATM principally binds to free DNA ends or DNA strand breaks (43), while ATR recognizes single-stranded regions of DNA common to ... cellular DNA, and that this damage leads to apoptosis Upon detection of DNA damage, DNA damage response proteins inhibit the cell cycle and are capable of inducing apoptosis if the DNA lesion ... with 5-aminoisoquinolinone The importance of DNA damage to NS1-induced apoptosis is shown by the abrogation of apoptosis in 93 the presence of inhibitors of DNA damage recognition enzymes Both the...

Ngày tải lên: 25/10/2012, 11:18

... nghiệm áp dụng qui trình PCR với DNA từ lông Dựa vào qui trình PCR nguồn DNA từ cơ, tiến hành xác định giới tính nguồn DNA từ lông Kết so sánh với kết PCR sử dụng nguồn DNA từ Thí nghiệm bố trí ... khô DNA kiểm tra thông qua ba tiêu Đó tỷ số OD, hàm lượng DNA thu hồi kết phân biệt giới tính phản ứng PCR (theo qui trình PCR tối ưu xác định) Bảng 3.2 Mức độ phơi khô cặn DNA Mức độ Biểu cặn DNA ... định giới tính ba giống bò dựa qui trình PCR tìm (4) So sánh hai mức độ làm khô DNA từ mẫu trình thu hồi DNA lên phản ứng PCR (5) So sánh hiệu PCR tiến hành DNA từ từ lông ba giống 3.2 THỜI GIAN...

Ngày tải lên: 29/10/2012, 14:13

... mặt thịt SUMMARY iv “The application of multiplex - PCR in detecting some virulent genes of E coli isolated from diarrhea faeces of bovine and swine, swabs of beef surface” The study was carried ... stx1, stx2, eae, ehxA and uid genes of isolated E coli from 27 diarrhea stools samples of bovines, calves, piglets, and swabs of the beef surface by multiplex - PCR There was the observing on each ... was chosen about – 10 separate colonies DNA of the colony groups and each separate colonies were extracted by the heat method The results of multiplex - PCR were showed that: (1) The colony group...

Ngày tải lên: 29/10/2012, 14:23

... phân lập multiplex - PCR - Việc phát gen độc lực E coli thực phản ứng multiplex – PCR với máy ln nhiệt (thermal cycler):  Multiplex – PCR1 : Phát gen eae, hly, stx1, stx2  Multiplex – PCR2 : Phát ... primer phản ứng Multiplex – PCR mơ tả Chamberlain năm 1988 kể từ multiplex – PCR ứng dụng nhiều lĩnh vực kiểm tra DNA (Protocol online) 2.2.2 Các giai đoạn phản ứng PCR Phản ứng PCR chuỗi gồm nhiều ... ion lần Thử IMViC(+/-;+;-;-) Ly trích DNA (DNA nhóm khuẩn lạc) Multiplex - PCR (-) Loại bỏ ống NA giữ gốc (+) Ly trích DNA riêng theo ống NA Multiplex- PCR Cạo tất khuẩn lạc chạm theo nhóm riêng:...

Ngày tải lên: 29/10/2012, 14:23

... pháp y hình Trong nghiên cứu genome, PCR ứng dụng nhân vô tính, multiplex PCR, cloning cDNA PCR đảo (inverse PCR) , recombinant PCR, Trong y khoa thú y, PCR sử dụng để chẩn đoán mầm bệnh virus, ... - 0,5 ml H2O cất khử ion lần Ly trích DNA (DNA nhóm khuẩn lạc) (-) Multiplex - PCR Loại bỏ ống NA giữ gốc (+) ) Ly trích DNA riêng theo ống NA Multiplex- PCR Sơ đồ 3.1 Qui trình phân lập, định ... độc lực E coli 3.3.4 Qui trình multiplex – PCR (1) Trình tự đoạn mồi sử dụng multiplex – PCR Bảng 3.1 Trình tự đoạn mồi sử dụng phản ứng multiplex - PCR 22 Kích cỡ DNA Mồi (primer) Trình tự oligonucleotide...

Ngày tải lên: 31/10/2012, 09:07

... Bảng 2.1 : Các loại marker DNA (Bùi Chí Bửu Nguyễn Thị Lang, 2005) Chỉ thị (marker) Tên đầy đủ RAPD Random Amplified Polymorphic DNA AP -PCR Arbitrary Primer -PCR DAF DNA Amplification Fingerprinting ... từ DNA mẫu Hình 2.3:Nguyên tắc phản ứng PCR 2.4.2 Thành phần vai trò chất phản ứng PCR  Mẫu DNA: Mẫu DNA đƣợc ly trích từ nhiều phận khác đối tƣợng nghiên cứu nhiều phƣơng pháp khác Đối với DNA ... ứng PCR khoảng 5-10 ng DNA khiết Nồng độ DNA xác định đƣợc nhờ đo OD (mật độ quang) độ dài sóng 260nm Khi DNA tinh khiết, lƣợng DNA đƣợc tính theo công thức : - OD260nm = 50 ng/ l (đối với DNA...

Ngày tải lên: 02/11/2012, 17:14

... sta, stb, vt2e genes of isolated E coli from 27 diarrhea stools samples of cattles, calves, piglets, and swabs of the beef surface by multiplex – PCR Observation had one group of pink colony on ... Những ứng dụng PCR .15 2.2.5.1 PCR sử dụng để nghiên cứu lượng DNA nhỏ .15 2.2.5.2 PCR sử dụng chẩn đoán lâm sàng .16 2.2.5.3 PCR dùng để khuếch đại RNA .16 vi 2.2.5.4 PCR sử dụng ... Nguyên lý phản ứng PCR 13 Hình 2.2 Chu kỳ nhiệt độ phản ứng PCR .14 Hình 3.1 Kết điện di sản phẩm multiplex - PCR1 26 Hình 3.2 Kết điện di sản phẩm multiplex - PCR2 26 SƠ ĐỒ...

Ngày tải lên: 15/11/2012, 09:28