核酸定序設施 DNA Sequencing

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圖譜優化-PeakTrace分析模式選擇說明
Instructions for Selecting PeakTrace Analysis Mode for Chromatogram Optimization

Peaktrace分析簡易說明:
核酸定序結果分析方式選擇說明

Explanation for Choosing DNA Sequencing Analysis Method

2024/03/22

核酸定序結果分析方式選擇說明:

(1) 定序實驗室提供原廠分析軟體 (KB Basecaller),適用於所有的定序結果分析。

(2) 我們也提供另一種分析軟體 PeakTrace Basecaller(自 2019 年 5 月起),可提升讀長並去除背景雜訊,並優化定序圖譜的解析度。

 

備註: 若需要這種額外分析,請在申請系統中勾選 Peaktrace 分析。

 

Peaktrace 分析建議適用於:

 

(1) 長片段 PCR 產物 > 500 bp 以及 Plasmid 的定序分析。

(2) 需要提高定序結果 850 bp 之後波峰解析度。

(3) 希望定序結果無任何小雜訊的客戶(請注意,這樣的修飾可能會忽略掉一些具有意義的小PEAK。

 

 

Peaktrace 分析不建議用在:

 

(1) 短片段 PCR 產物 < 500 bp(500 bp 之前的解析度已經很好,無需特別優化)。

(2) 基因編輯類實驗或預期定序結果出現 multiple peaks 時,如 CRISPR 等等。

(3) 基因型分析,如 SNP 或突變位點確認等等(為了避免 minor peak 被忽略)。

(4) 失敗的定序反應。

 

備註:

(1) Peaktrace 分析結果顯著改善僅適用於品質與濃度均達到規定標準的定序樣品。

(2) 在 Bio201 申請系統完善建立之前,選擇 Peaktrace 分析的客戶若對結果有疑慮,可向定序中心索取原廠分析結果。

(3) 未來,選擇 Peaktrace 分析時,我們將同步提供原廠分析與 Peaktrace 分析結果。

備註: 上述說明在協助您選擇最適合您特定定序需求的分析方法。如果您有任何疑問或需要進一步的說明,請隨時與我們核心設施聯繫。2652-3924

Explanation for Choosing DNA Sequencing Analysis Method:

(1) Our core facility provide the manufacturer's analysis software (KB Basecaller) which is suitable for all sequencing result analyses.

Note: Clients do not need to specify this on the order; we will prioritize providing results from the manufacturer's analysis.

 

(2) We also offer the PeakTrace Basecaller (since May 2019), which improves read lengths, removes background noise, and optimizes the resolution of sequencing chromatograms.

Note: If you require this additional analysis, please select the use of PeakTrace in the the order.

 

PeakTrace analysis is more suitable for:

(1) Sequencing of long PCR products (> 800 bp) and Plasmids.

(2) Customers needing improved resolution of peak chromatograms beyond 850 bp.

(3) Customers who prefer sequencing results without any minor noise (please note that this modification may overlook some meaningful small peaks).

 

Not recommended for:

(1) Short PCR products (< 500 bp, as resolution before 500 bp is well-defined).

(2) Gene editing experiments or situations with multiple peaks expected in sequencing results (e.g., CRISPR, etc.).

(3) Genotyping experiments, such as SNP or mutation site confirmation (to avoid ignoring minor peaks).

(4) Failed sequencing reactions.

 

Note:

(1) Significant improvements from PeakTrace analysis require samples to match the required concentration and quality standards..

(2) Before the Bio201 application system is fully established, customers choosing PeakTrace analysis can request the original manufacturer's analysis results from the sequencing core if they have concerns.

(3) In the future, when selecting PeakTrace analysis, we will concurrently provide both the manufacturer's analysis and PeakTrace analysis results.

 Note: The above information is provided to ensure transparency and assist you in choosing the most suitable analysis method for your specific sequencing needs. If you have any questions or need further clarification, please don't hesitate to reach out to our core.