30
2022．05
快速上手！最簡單的DNA萃取方法
提取基因組DNA (genomic DNA, gDNA)是一種常規的分子生物學實驗室技術。gDNA包含生物體的所有遺傳、染色體物質，它編碼生命所必需的所有成分，研究單個基因或對整個基因組進行測序等等實驗[1]，是大規模流行病學研究中至關重要的第一步[2, 3]。
隨著這個流感、病毒快速來襲的時代，各種疾病研究中，對於gDNA分析的需求不斷增加，增加了對大量gDNA提取的需求，如何快速有效的萃取高質量的gDNA並且進行下游的分析至關重要。提取gDNA的樣本形式與應用非常多元，如：收集口腔粘膜刮樣，用於檢測口腔生菌數與口腔癌誘發因子；進行深喉痰液與鼻腔採樣，用於檢測呼吸道病毒；從血液樣本中提取gDNA來檢測和監測癌症患者的疾病等[4-6]。
然而傳統的提取gDNA的各種方法一直存在著許多缺點，包括：程序繁雜、時間長、產量低、成本高、品質受損、使用有毒有機溶劑等，並且僅適用於某些類型的組織[3, 7-11]。
圖爾思為快速提取DNA提供了簡單、省時和經濟的方法：EX-TOOL Quick DNA Extraction Solution (TTJ-QS100)，此產品簡化許多操作步驟，比傳統的提取方法減少更多步驟和時間，不需使用任何有毒化學品或任何額外的材料，只需要簡單的混和與加熱處理，且只需要3 - 6分鐘即可有效萃取gDNA；實驗證明其所製備的樣本適用於PCR或是qPCR等分子檢測方法學，每批驗證品質穩定，適合分子檢測開發與高通量使用。EX-TOOL Quick DNA Extraction Solution適用於各種樣本，包含各種動物組織、細胞和全血。
隨著這個流感、病毒快速來襲的時代，各種疾病研究中，對於gDNA分析的需求不斷增加，增加了對大量gDNA提取的需求，如何快速有效的萃取高質量的gDNA並且進行下游的分析至關重要。提取gDNA的樣本形式與應用非常多元，如：收集口腔粘膜刮樣，用於檢測口腔生菌數與口腔癌誘發因子；進行深喉痰液與鼻腔採樣，用於檢測呼吸道病毒；從血液樣本中提取gDNA來檢測和監測癌症患者的疾病等[4-6]。
然而傳統的提取gDNA的各種方法一直存在著許多缺點，包括：程序繁雜、時間長、產量低、成本高、品質受損、使用有毒有機溶劑等，並且僅適用於某些類型的組織[3, 7-11]。
圖爾思為快速提取DNA提供了簡單、省時和經濟的方法：EX-TOOL Quick DNA Extraction Solution (TTJ-QS100)，此產品簡化許多操作步驟，比傳統的提取方法減少更多步驟和時間，不需使用任何有毒化學品或任何額外的材料，只需要簡單的混和與加熱處理，且只需要3 - 6分鐘即可有效萃取gDNA；實驗證明其所製備的樣本適用於PCR或是qPCR等分子檢測方法學，每批驗證品質穩定，適合分子檢測開發與高通量使用。EX-TOOL Quick DNA Extraction Solution適用於各種樣本，包含各種動物組織、細胞和全血。
- 實驗流程
- 產品特色
超簡單：僅需加熱與震盪，即可萃取核酸
省時：實驗證實6分鐘即可有效萃取
應用性廣：實驗證實後端可用於PCR與qPCR，使用方法簡單，極適合高通量樣本處理與分析
- 實驗數據
EX-TOOL Quick DNA Extraction Solution經過驗證，整個實驗只需要3 - 6分鐘即可有效萃取DNA (圖一)。使用EX-TOOL Quick DNA Extraction Solution備置的gDNA樣本相當穩定，儲存穩定性測試，結果顯示含EDTA的人類血液樣本以EX-TOOL Quick DNA Extraction Solution提取，儲存14天後使用於PCR，其結果依然與未存放之樣本一樣(圖二)。另外，亦可使用於各種動物組織樣本，如：常見的模式動物，小鼠的脾臟、心臟和血液(圖三)，以及特殊的鸚鵡的血液和羽毛 (羽根)，從中提取發生於鸚鵡常見的病毒感染 (圖四、五)。
- Figure 1：Genomic DNA of human blood were prepared with EDTA by EX-TOOL Quick DNA Extraction Solution (TTJ-QS01). The samples were extracted with variety of incubation time and then used for PCR amplification using TOOLS 2X SuperGreen PCR Master Mix (TTC-PA31-10) to examine the extraction efficiency by electrophoresis (100V, 1% TAE agarose gel prepared with TOOLS DNA View (TT-DNA01, BIOTOOLS) in 1X TAE buffer (TT-50TA, TOOLS) for 30 minutes. NTC: no template control. M: TOOLS 1kb Ladder (TMG1kb, BIOTOOLS).Figure 1：Genomic DNA of human blood were prepared with EDTA by EX-TOOL Quick DNA Extraction Solution (TTJ-QS01). The samples were extracted with variety of incubation time and then used for PCR amplification using TOOLS 2X SuperGreen PCR Master Mix (TTC-PA31-10) to examine the extraction efficiency by electrophoresis (100V, 1% TAE agarose gel prepared with TOOLS DNA View (TT-DNA01, BIOTOOLS) in 1X TAE buffer (TT-50TA, TOOLS) for 30 minutes. NTC: no template control. M: TOOLS 1kb Ladder (TMG1kb, BIOTOOLS).
- Figure 2：Genomic DNA from human blood with EDTA were prepared with EX-TOOL Quick DNA Extraction Solution (TTJ-QS01). The extracted samples further were taken to perform PCR amplification using TOOLS 2X SuperGreen PCR Master Mix (TTC-PA31-10) immediately or after stored for 3, 7, 14 days, The PCR products of SOX21, 237 bp, in every sample were then examined with electrophoresis (100V, 1% TAE agarose gel prepared with TOOLS DNA View (TT-DNA01, BIOTOOLS) in 1X TAE buffer (TT-50TA, TOOLS) for 30 minutes. DNA size was referenced with DNA Ladder. NTC: no template control.
- Figure 3：Genomic DNA from mouse spleen, heart, kidney, blood are prepared with EX-TOOL Quick DNA Extraction Solution (TTJ-QS01). The extracted samples were then used for PCR amplification using TOOLS 2X SuperGreen PCR Master Mix (TTC-PA31-10) examine the extraction efficiency with electrophoresis (100V, 1% TAE agarose gel prepared with TOOLS DNA View (TT-DNA01, BIOTOOLS) in 1X TAE buffer (TT-50TA, TOOLS) for 30 minutes. DNA size was referenced with TOOLS 1kb Ladder (TMG1kb, BIOTOOLS). PC-1: mouse cell cDNA diluted with TTJ-QS01, PC-2: mouse cell cDNA diluted with ddH2O, NTC: no template control.
- Figure 4：Genomic DNA from whole blood of parrot was prepared with EX-TOOL Quick DNA Extraction Solution (TTJ-QS01). The extracted sample was then used for PCR amplification and electrophoresis to examine the extraction efficiency. The amplicon size was referenced as 500 bp with DNA ladder (lane M).
- Figure 5：Genomic DNA from feather tip was prepared with EX-TOOL Quick DNA Extraction Solution (TTJ-QS01). The extracted sample was then used for PCR amplification and electrophoresis to examine the extraction efficiency. The amplicon size was referenced as 500 bp with DNA ladder (lane M).
Reference
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