1 / 17

核酸定序核心 DNA S equencing C ore

研發分處第一共同研究室 技術服務說明會. 核酸定序核心 DNA S equencing C ore. Applications overview. • De novo sequencing of genomes • Detection of variants (SNPs) and mutations • Confirmation of clone constructs • Detection of methylation events • Gene expression studies • Detection of copy number variation.

elaine-hudson
Download Presentation

核酸定序核心 DNA S equencing C ore

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. 研發分處第一共同研究室 技術服務說明會 核酸定序核心DNA Sequencing Core

  2. Applications overview • De novo sequencing of genomes • Detection of variants (SNPs) and mutations • Confirmation of clone constructs • Detection of methylation events • Gene expression studies • Detection of copy number variation

  3. Automated DNA Sequencing Workflow • PCR products • Plasmid DNA • Genomic DNA • Bacterial artificial chromosomes (BACs) • Yeast artificial chromosomes (YACs) • Cosmids DNA Template preparation Cycle sequencing Applied Biosystems Cycle Sequencing Kits: • BigDye Terminator v3.1 kit • dGTP BigDye Terminator v3.0 kit Extension Product Purification Purify extension products using Ethanol precipitation 3730xl analyzer Applied Biosystems instruments: Capillary Electrophoresis Capillary Electrophoresis Output: *.ab1 file Applied Biosystems Sequencing Analysis Software V5.4 Data analysis

  4. DNA Quality Contamination Potential contaminants include: • Proteins • RNA • Chromosomal DNA • Excess PCR primers, dNTPs, enzyme, and buffer components (from a PCR amplification used to generate the sequencingtemplate) • Residual salts • Residual organic chemicals such as phenol, chloroform, and ethanol • Residual detergents Determining DNA Quality • Agarose gel electrophoresis • Spectrophotometry The A260/A280 ratio should be 1.7 to 1.9.

  5. DNA Quantity PCR product請以電泳分析比對marker確認濃度。 • Primer quantity 6 pmol (6μM*1μl) • 一件樣品＝ 1 DNA template＋1 端primer (自行提供)或 • 一件樣品＝1 DNA template (使用用本實驗室的primer) • (在200 ul微量離心管，將DNA template與primer混合或template only，補ddH2O至最後體積6μl)

  6. 系統連結位置 轉自:計中-傅潔瑩小姐

  7. 系統使用頁面 轉自:計中-傅潔瑩小姐

  8. 系統登入 轉自:計中-傅潔瑩小姐

  9. PI授權管理頁面 轉自:計中-傅潔瑩小姐

  10. PI-帳務月報表 轉自:計中-傅潔瑩小姐

  11. PI-系統回報每日新增預約單 轉自:計中-傅潔瑩小姐

  12. 一般使用者-系統服務行事曆 轉自:計中-傅潔瑩小姐

  13. 一般使用者-預約單填寫 轉自:計中-傅潔瑩小姐

  14. 一般使用者-預約記錄查詢 轉自:計中-傅潔瑩小姐

  15. 一般使用者-實驗結果下載 轉自:計中-傅潔瑩小姐

  16. 收費流程 • 每個序列分析反應醫學院院區之收費為新台幣100元整，非醫學院院區為新台幣150元整。收費採月結方式。 • 分析試驗必須先上網預約，再送樣品。如只收到樣品而未收到預約單，視同未申請，本實驗室也不負責保管此樣品。如只收到預約單而未收到樣品，視同已申請，須付費。 • 每個月月初管理人員會依實驗室(或計畫)主持人為對象整理上一個月之使用額度，統計總額，送交醫學院出納股辦理預開收據。 • 管理人員依實驗室(或計畫)主持人之使用量，填妥繳費單(月結) 表格暨預開收據通知使用人辦理會計核帳之程序。 • 繳費單(月結) 表格經使用人確認金額無誤簽收後，把繳費單(月結)及紅色收據聯交負責實驗室計劃的帳務人員報帳，開立黏存單繳費。 • 使用者若選擇以現金支付，請持繳費單(月結) 及現金至醫學院出納股直接繳費。

  17. 感謝聆聽 負責人: 網路系統支援: 李芳仁 主任 計資中心-傅潔瑩小姐 鄧述諄 副主任 林泰元 老師聯絡人:階孟書

More Related