200535244 玖、發明說明： 【發明所屬之技術領域】 本發明係關於一種能提高聚合酶鏈鎖反應放大效果的 聚合酶鏈鎖反應溶液及其應用。 【先前技術】 現今在處理石化油〉亏的污染場址時，生物復育法 (bioremediation)已經廣受重視。生物復育法（Bioremediaticm) 係利用自然程序清除環境中的有害物質。存活於土壤與地 下水中的微生物將特定的有害物質，例如汽油與漏油中的 有害物質’當作能量來源，經由生化代謝反應，有害物質 最終轉變成為水與無害的生理代謝物。為了讓生物復育順 利進行，必需保持微生物健康與活性，生物復育技術是與 微生物的成長、增殖息息相關的，若要有最佳的去除污染 物效果，就要提供微生物最適的環境條件。特定的生物復 育技術決定於以下幾個因素：已存在的微生物相、微生物 能分解的污染物種類以及存活的環境狀況。因此，在進行 生物復育的操作之前，土壤中的微生物相是我們所必須了 解的’若欲了解可分解石化油類化合物之微生物之組成以 及其喜好之環境，研究土壤中微生物社會結構是必需的。 除了傳統分菌培養法之外，利用分子生物的方法萃取DNA 以及聚合酶鏈鎖反應（polymerase chain reaction，PCR)放大 反應常為研究環境中微生物的社會結構時的第一步驟。 聚合酶鏈鎖反應（poly merase chain reaction，PCR)是目 前最常應用於放大DNA的一個技術，它可使DNA在微量 試管中擴增至1〇6倍以上。PCR的原理十分簡單，首先在 欲擴增的DNA片段兩端分別設計一個前置引子（f〇rward 200535244 primer)和反置引子（reverse primer)使之與已變性的單股目 標DNA緩冷配對（annealing)後，利用DNA聚合酵素(DNA polymerase)以目標DNA的兩股分別做為模板(template)來 合成新的DNA股。現行的操作技術一般是將DNA模板加 入含包含引子、聚合酶、dNTP及緩衝液的反應溶液後，在 不同溫度進行三個步驟來達成放大DNA的效果：（1)變性 反應（denaturation)，使DNA的兩股分離；（2)緩冷配對反應 (annealing)，使引子與目標DNA配對，·（3)延長反應 (enlogation)，合成新的DNA股。理論上，當PCR反應進 行η次時，DNA的分子數可達到2n個分子數。 然而，聚合酶鏈鎖反應的效果往往會受到萃取出的 DNA之中含有的抑制物質所影響，進而限制了其可用性。 已知有許多會抑制聚合酶放大反應的物質，如血液中的血 基質（Heme)及其衍生物，以及眾多污染環境樣本中所含有 的多環化合物，像腐植酸等皆會干擾聚合酶鏈鎖反應的進 行。因此，當此PCR技術使用在環境樣本DNA的放大時， 由於環境樣本所含的不明物質偏多且在萃取DNA時常無 法完全去除’所以卒取之出DNA中常帶有一些會抑制PCR 反應進行之物質，如腐植酸、某些環狀化合物或是苯酚類 荨專’造成PCR反應然法有效完成，因而無法順利取得放 大產物。現有的DNA萃取技術雖然能萃取出大部分實驗樣 本的DNA，但是並無法完全去除包含在其中的抑制物質， 為了得到可進行聚合梅放大反應的DNA樣本，通常都需要 透過純化的方式，但是隨著純化的步驟愈多，所流失的 DNA樣本相對的增加，進而影響到實驗結果的正確性。因 此’尋求另一種途徑來得到適用的DNA樣本而不流失過多 200535244 的DNA是必須的。 【發明内容】 針對習知聚合酶鏈鎖反應的問題，本發明係關於一種 聚合酶鏈鎖反應溶液，包含前置引子、反置引子、募核苷 酸、DNA聚合晦、緩衝液及水，其特徵係為：前述溶液進 一步包含牛血清白蛋白（bovine serum albumin，BSA)，其中 該牛血清白蛋白之含量係為5%-25%(w/w)。 本發明之另一目的係提供一種放大環境樣本中DNA的 方法，包含： (a) 洗去環境樣本中的游離離子； (b) 離心將雜質沉澱，取得懸浮菌液； (c) 將細胞打破，取得游離之DNA ; (d) 將DNA析出，獲得環境樣本中存在之DNA ; (e) 將步驟（d)獲得之DNA加入聚合酶鏈鎖反應溶液 中，其中聚合酶鏈鎖反應溶液包含前置引子、反 置引子、募核苷酸、DNA聚合晦、緩衝液、牛 血清白蛋白及水，其中反應溶液中牛血清白蛋白 之含量係為5%-25%(w/w);及 (f) 進行聚合酶鏈鎖反應，獲得放大之環境DNA樣 本0 本發明係添加牛血清白蛋白（BSA)至PCR反應溶液 中，以加強PCR的放大效果。BSA具有保護酵素的功用， 能保護酵素使其不失去作用能力，降低樣本中所含不易去 除之抑制物質對聚合酶的影響，增加聚合酶靈敏度及作用 能力，進而增進PCR放大效果。本發明特別適用於分析環 200535244 境樣本中的微生物，可有效解決樣本中抑制物質影響PCR 放大效果的問題。 【實施方式】 本發明係關於一種聚合酶鏈鎖反應溶液，包含前置引 子、反置引子、寡核苷酸、DNA聚合酶、緩衝液及水，其 特徵係為：前述溶液進一步包含牛血清白蛋白，其中該牛 血清白蛋白之含量係為5%-25%，較佳為10%〜20%(w/w)， 最佳為20%(w/w)。 本發明之聚合酶鏈鎖反應溶液，其中聚合酶較佳係為 Taq DNA聚合酶。前述之寡核苷酸係為去氧核醣核酸三磷 酸（Deoxyribonucleoside Triphosphate，dNTP)混合物，包含 dATP、dCTP、dGTP 與 dTTP 〇 本發明提供之聚合酶鏈鎖反應溶液，係可有效解決環 境樣本中經常包含未知抑制物造成PCR放大效果不佳的問 題’能有效自環境中萃取DNA並經PCR放大DNA樣本。 因此，本發明之另一目的即提供一種放大環境樣本中DNA 的方法’其步驟包含：首先，洗去環境樣本中的游離離子； 接著’離心將雜質沉澱，取得懸浮菌液；然後打破細胞， 將蛋白質以及懸浮的胞器析出，取得游離之DNA ; DNA純 化析出後’加入本發明之聚合酶鏈鎖反應溶液中，其中聚 合酶鏈鎖反應溶液包含前置引子、反置引子、寡核苷酸、 DNA聚合酶、緩衝液、牛血清白蛋白及水，其中反應溶液 中牛血清白蛋白之含量係控制在5%-25%;最後設定適當之 變性反應、緩冷配對反應與延伸反應溫度，進行聚合酶鏈 鎖反應’最終可獲得放大數倍之環境DNA樣本。 本發明提供之聚合酶鏈鎖反應溶液，特別適合用於油 200535244 污、=或是腐植酸含量高之環境樣本PCR放大程序。前述反 應/合液中所含之聚合酶例如，但不限於：Taq聚合酶。 、本發明之實施例係詳細描述於下，然而下列實施例係用 於了解本發明之優點，並非用於限制本發明之申請專利範 圍。 境樣本DNA之萃取 ，本來源：以加油站土壤為樣本，萃取樣本中的dna。 萃取環境樣本中DNA的方法如下：首先洗去環境樣 本中的游離離子；然後以超高速離心將雜質沉澱，取得懸 浮菌液，接著以磁珠將細胞打破，以苯酚/三氣甲烷/生長 素〇引讦_3_乙酸）將蛋白質以及懸浮的胞器析出，取得游離 之DNA ;最後以100%酒精將DNA析出，獲得環境樣本 中總菌群之DNA。 實施例1 在PCR管中加入實施例1萃取之土壤樣本中總菌群之 DNA及PCR反應溶液。PCR反應溶液組成如表一所示， 包含 2// 1 PCR 10x buffer(20mM Tris-HCl，l〇〇mM KC1， O.lmM EDTA，ImM DTT，0.5% Tween 20，〇·5〇/0 Nonidet P-40，50% Glycerol)、1.6 " 1 dNTP mix、〇·4 // 1 前置引子、 0.4//1 反置引子、0.1//1 Taq 聚合酶、11 // 1 h20、及 4 // 1 BSA。 設定PCR操作條件：（a)變性反應：95°C，3分鐘。（b)28 循環·· 95°C，1 分 30 秒；52°C，〇 分 45 秒；72°C，2 分 鐘。（c)延伸反應：72°C，3分鐘。 200535244 將PCR管置入反應器中進行反應，然後取10/ζι之 PCR產物進行電泳分析，結果如第一圖所示。 表一 PCR 反應溶液組成（含BSA) 10X buffer :2β\ dNTP mix :1.6//1 前置引子 :0.4// 1 反置引紫 :0.4 β\ Taq聚合酶 :0.1 β 1 H20 :ΙΙβΙ BSA :4β\ Total :20 μ\ 比較實施例1 在PCR管中加入實施例1萃取之土壤樣本中總菌群 之DNA及PCR反應溶液。PCR反應溶液組成如表二所 示，為傳統進行PCR反應之溶液組成：包含2 /z 1 PCR 1 Ox buffer(20mM Tris-HCl，100mM KC1，O.lmM EDTA，ImM DTT，0.5% Tween 20，0.5% Nonidet P-40，50〇/〇 Glycerol)、 1.6 # 1 dNTP mix、0.4// 1 前置引子、0.4 # 1 反置引子、 0.1 // 1 Taq 聚合酶及 15/z 1 H20。 設定PCR反應條件：（a)變性反應：95°C，3分鐘。（b)28 循環：95t：，1 分 30 秒；52°C，〇 分 45 秒；72°C , 2 分 鐘。（c)延伸反應：72°C，3分鐘。 將PCR管置入反應器中進行反應，然後取1 〇 # 1之 200535244 PCR產物進行電泳分析，結果如第二圖所示 表二：PCR反應溶液組成（無BSA) 2 β 1 1.6 β\ 0.4//1 0.4//1 0.1 β 1 15//1 20//1 10X buffer dNTP mix 前置引子 反置引子 Taq聚合酶 H20_ 總量 比較結果 第一圖係使用本發明之包含BSA的PCR反應溶液放 大DNA後的PCR產物電泳圖。第二圖係使用傳統未含 BSA的PCR反應溶液放大DNA後的PCR產物電泳圖。 比較第一圖及第二圖之結果，可發現第一圖之編號6及7 之樣本有明顯亮帶，第二圖則完全無任何明顯之亮帶。此 一結果顯示實施例一的PCR產物量高於實施例二之產 量，換言之，即在PCR反應溶液組成中添加bsA後能獲 得較高之PCR產量，使低量或萃取不易之〇ΝΑ樣本能有 效經PCR放大進行進一步檢測。 實施例2 將實施例1萃取之DNA稀釋1 〇倍。在pcr管中加 入前述稀釋10倍之土壤樣本中總菌群之DNA及表一之 PCR反應溶液。設定PCR反應溶液，將pcr管置入反應 200535244 器中進行反應，然後取10// 1之PCR產物進行電泳分析， 結果如第三圖所示。其中PCR之反應條件及電泳分析方 法與實施例1相同。 比較實施例2 將實施例1萃取之DNA稀釋1 〇倍。在PCR管中加入 前述稀釋1 〇倍之土壤樣本中總菌群之DNA及表二之PCR 反應溶液。設定PCR反應溶液，將pCR管置入反應器中 進行反應，然後取10/zl之PCR產物進行電泳分析，結果 如第三圖所示。其中PCR之反應條件及電泳分析方法與 實施例1相同。 比較結果 第三圖係使用本發明之包含BSA的pCR反應溶液放 大稀釋10倍之DNA後的PCR產物電泳圖。第四圖係使 用傳統未含BSA的PCR反應溶液放大稀釋1〇倍之 後的PCR產物電泳圖。比較第三圖及第四圖之結果，可 發現第三圖之編號1、2、3、4、6、7、8及9之樣本有明 顯亮帶，第二圖則僅2、3、6及7之樣本有明顯亮帶。此 一結果顯示實施例三的PCR產物量高於實施例肆之產 量’換言之，即在PCR反應溶液組成中添加BSA後能獲 得較高之PCR產量，使低量或萃取不易之DNA樣本能有 效經PCR放大進行進一步檢測。 其他實施態樣 所有本說明書所揭露之特徵皆可以任何形式與其他方 12 200535244 法合併使m明書中所揭露之特徵可以具有相同、相 等或相似目的的特徵所取代。因此，&了明確強調的部分 之外’所有本說明書中揭露的特徵僅為眾多相等或相似特 徵中的一個實施例。 根據本說明書所揭露的内容，任何一位熟習本技術領 域的人士可輕易明確瞭解本發明之重要特色在不脫離本 發明^精神與範圍下，係可對本發明做不同的改變與修飾 使其符合不同的使用目的與情況。因此，其他實施態樣亦 包含在下列申請專利範圍内。 【圖式簡單說明】 第一圖係為未稀釋之土壤DNA樣本PCR產物電泳圖，其中 PCR反應溶液係為本發明之包含bsa之反應溶液。 第二圖係為未稀釋之土壤DNA樣本PCR產物電泳圖，其中 PCR反應溶液係為傳統未包含bsa之反應溶液。 第三圖係為稀釋10倍之土壤DNA樣本PCR產物電泳圖， 其中PCR反應溶液係為本發明之包含bsa之反應溶液。 第四圖係為稀釋10倍之土壤DNA樣本PCR產物電泳圖， 其中PCR反應溶液係為傳統未包含BSA之反應溶液。 13200535244 (1) Description of the invention: [Technical field to which the invention belongs] The present invention relates to a polymerase chain reaction solution capable of improving the amplification effect of the polymerase chain reaction and its application. [Previous Technology] Nowadays, bioremediation has been paid much attention when dealing with contaminated sites with petrochemical oil> deficiency. Bioremediaticm uses natural procedures to remove harmful substances from the environment. Microorganisms living in soil and underground water use specific harmful substances, such as the harmful substances in gasoline and oil spills, as energy sources. Through the biochemical metabolic reaction, the harmful substances are finally transformed into water and harmless physiological metabolites. In order for biological rejuvenation to proceed smoothly, it is necessary to maintain the health and activity of microorganisms. Biological rejuvenation technology is closely related to the growth and proliferation of microorganisms. To have the best effect of removing pollutants, it is necessary to provide the most suitable environmental conditions for microorganisms. The specific bioremediation technology depends on the following factors: the existing microbial phase, the types of pollutants that the microorganism can decompose, and the environmental conditions in which it survives. Therefore, before the operation of biological rehabilitation, the microbial phase in the soil is what we must understand. 'If you want to understand the composition of the microorganisms that can decompose petrochemical oil compounds and their preferred environment, it is necessary to study the social structure of microorganisms in the soil. of. In addition to traditional bacterial culture methods, the use of molecular biology methods to extract DNA and polymerase chain reaction (PCR) amplification reactions are often the first steps in studying the social structure of microorganisms in the environment. Polymerase chain reaction (PCR) is currently the most commonly used technique for amplifying DNA, and it can amplify DNA by more than 106 times in micro-tubes. The principle of PCR is very simple. Firstly, design a pre primer (fowler 200535244 primer) and a reverse primer on the two ends of the DNA fragment to be amplified to slowly pair with the denatured single-stranded target DNA. (Annealing), using DNA polymerase to synthesize new DNA strands using the two strands of the target DNA as templates, respectively. The current operation technique is generally to add a DNA template to a reaction solution containing a primer, a polymerase, dNTP and a buffer solution, and then perform three steps at different temperatures to achieve the effect of amplifying DNA: (1) denaturation, so that The two strands of DNA are separated; (2) slow cooling pairing (annealing), pairing the primer with the target DNA, and (3) prolonging the reaction (synthesis) to synthesize a new DNA strand. Theoretically, when the PCR reaction is performed n times, the number of DNA molecules can reach 2n. However, the effect of the polymerase chain reaction is often affected by the inhibitory substances contained in the extracted DNA, which limits its usability. Many substances known to inhibit the polymerase amplification reaction, such as blood matrix (Heme) and its derivatives in blood, and polycyclic compounds contained in many environmentally polluted samples, such as humic acid, can interfere with the polymerase chain The lock reaction proceeds. Therefore, when this PCR technology is used to amplify DNA in environmental samples, because environmental samples contain too many unknown substances and cannot be completely removed when extracting DNA, so the DNA that is taken out often contains some that will inhibit the PCR reaction. Substances such as humic acid, certain cyclic compounds, or phenolic compounds have caused the PCR reaction to be effectively completed, and thus cannot successfully obtain amplified products. Although the existing DNA extraction technology can extract the DNA of most of the experimental samples, it cannot completely remove the inhibitory substances contained in it. In order to obtain DNA samples that can be amplified by polymer plums, purification methods are usually required. The more purification steps there are, the more DNA samples are lost, which will affect the accuracy of the experimental results. Therefore, it is necessary to seek another way to obtain a suitable DNA sample without losing too much 200535244 DNA. [Summary of the Invention] To solve the problem of the conventional polymerase chain reaction, the present invention relates to a polymerase chain reaction solution, which includes a pre-primer, an inverted primer, nucleotides, DNA polymerization, buffer, and water. It is characterized in that the aforementioned solution further comprises bovine serum albumin (BSA), wherein the content of the bovine serum albumin is 5% -25% (w / w). Another object of the present invention is to provide a method for amplifying DNA in an environmental sample, comprising: (a) washing away free ions in the environmental sample; (b) precipitating impurities by centrifugation to obtain a suspension bacteria solution; (c) breaking cells (D) Precipitate the DNA to obtain the DNA present in the environmental sample; (e) Add the DNA obtained in step (d) to the polymerase chain reaction solution, where the polymerase chain reaction solution contains Placement of primers, reverse primers, nucleotide collection, DNA polymerization, buffer, bovine serum albumin and water, wherein the content of bovine serum albumin in the reaction solution is 5% -25% (w / w); and (f) Perform a polymerase chain reaction to obtain amplified environmental DNA samples. 0 The present invention adds bovine serum albumin (BSA) to the PCR reaction solution to enhance the amplification effect of PCR. BSA has the function of protecting enzymes. It can protect enzymes from losing their ability to reduce the effect of inhibitors contained in the sample on the polymerase, increase the sensitivity and ability of the polymerase, and thereby improve the PCR amplification effect. The invention is particularly suitable for analyzing microorganisms in environmental samples of 200535244 environment, and can effectively solve the problem that the inhibitory substances in the sample affect the PCR amplification effect. [Embodiment] The present invention relates to a polymerase chain reaction solution, which comprises a pre-primer, an inverted primer, an oligonucleotide, a DNA polymerase, a buffer solution and water, and is characterized in that the aforementioned solution further comprises bovine serum Albumin, wherein the content of the bovine serum albumin is 5% -25%, preferably 10% -20% (w / w), and most preferably 20% (w / w). The polymerase chain reaction solution of the present invention, wherein the polymerase is preferably Taq DNA polymerase. The aforementioned oligonucleotide is a mixture of Deoxyribonucleoside Triphosphate (dNTP), which includes dATP, dCTP, dGTP and dTTP. The polymerase chain reaction solution provided by the present invention can effectively solve the problems in environmental samples. The problem of poor PCR amplification due to the inclusion of unknown inhibitors can effectively extract DNA from the environment and amplify DNA samples by PCR. Therefore, another object of the present invention is to provide a method for amplifying DNA in an environmental sample. Its steps include: first, washing away free ions in the environmental sample; and then 'centrifuging to precipitate impurities to obtain a suspension bacteria solution; and then breaking the cells, The protein and suspended organs are separated to obtain free DNA; after the DNA is purified and precipitated, it is added to the polymerase chain reaction solution of the present invention, wherein the polymerase chain reaction solution includes a preprimer, an inverted primer, and an oligonucleoside. Acid, DNA polymerase, buffer, bovine serum albumin, and water, where the content of bovine serum albumin in the reaction solution is controlled at 5% -25%; finally, the appropriate denaturation reaction, slow cooling pairing reaction and extension reaction temperature The polymerase chain reaction is performed to finally obtain an environmental DNA sample at multiple magnifications. The polymerase chain reaction solution provided by the present invention is particularly suitable for the PCR amplification procedure of environmental samples with high oil content and high humic acid content. The polymerase contained in the aforementioned reaction / synthesis liquid is, for example, but not limited to, Taq polymerase. The embodiments of the present invention are described in detail below. However, the following embodiments are used to understand the advantages of the present invention and are not intended to limit the scope of patent application of the present invention. Extraction of DNA from environmental samples. Source: DNA from the gas station was used as a sample to extract DNA from the sample. The method of extracting DNA from environmental samples is as follows: first, the free ions in the environmental samples are washed away; then the impurities are precipitated by ultra-high speed centrifugation to obtain a suspension bacteria solution, and then the cells are broken by magnetic beads, and phenol / three gas methane / auxin 〇 引 讦 _3_acetic acid) to separate proteins and suspended organelles to obtain free DNA; finally, 100% alcohol to isolate the DNA to obtain the DNA of the total flora in environmental samples. Example 1 The DNA of the total flora in the soil sample extracted in Example 1 and the PCR reaction solution were added to a PCR tube. The composition of the PCR reaction solution is shown in Table 1. It contains 2 // 1 PCR 10x buffer (20mM Tris-HCl, 100mM KC1, O.lmM EDTA, ImM DTT, 0.5% Tween 20, 0.5 / 50 Nonidet P-40, 50% Glycerol), 1.6 " 1 dNTP mix, 0 · 4 // 1 pre-primer, 0.4 // 1 reverse primer, 0.1 // 1 Taq polymerase, 11 // 1 h20, and 4 // 1 BSA. Set PCR operating conditions: (a) Denaturation reaction: 95 ° C, 3 minutes. (B) 28 cycles · 95 ° C, 1 minute and 30 seconds; 52 ° C, 0 minutes and 45 seconds; 72 ° C, 2 minutes. (C) Extension reaction: 72 ° C, 3 minutes. 200535244 Put the PCR tube into the reactor for reaction, and then take the 10 / ζι PCR product for electrophoresis analysis. The result is shown in the first figure. Table 1 Composition of PCR reaction solution (including BSA) 10X buffer: 2β \ dNTP mix: 1.6 // 1 Pre-primer: 0.4 // 1 Inverted primer: 0.4 β \ Taq polymerase: 0.1 β 1 H20: ΙΙβΙ BSA: 4β \ Total: 20 μ \ Comparative Example 1 Add the DNA and PCR reaction solution of the total flora in the soil sample extracted in Example 1 to the PCR tube. The composition of the PCR reaction solution is shown in Table 2. It is the composition of the traditional PCR reaction solution: it contains 2 / z 1 PCR 1 Ox buffer (20mM Tris-HCl, 100mM KC1, O.lmM EDTA, ImM DTT, 0.5% Tween 20, 0.5% Nonidet P-40, 50〇 / 〇Glycerol), 1.6 # 1 dNTP mix, 0.4 // 1 pre-primer, 0.4 # 1 reverse primer, 0.1 // 1 Taq polymerase and 15 / z 1 H20. Set PCR reaction conditions: (a) Denaturation reaction: 95 ° C, 3 minutes. (B) 28 cycles: 95t: 1 minute and 30 seconds; 52 ° C, 0 minutes and 45 seconds; 72 ° C, 2 minutes. (C) Extension reaction: 72 ° C, 3 minutes. Place the PCR tube in the reactor for the reaction, and then take the 1035 # 200535244 PCR product for electrophoresis analysis. The results are shown in the second figure. Table 2: Composition of the PCR reaction solution (without BSA) 2 β 1 1.6 β \ 0.4 // 1 0.4 // 1 0.1 β 1 15 // 1 20 // 1 10X buffer dNTP mix Pre-primer reverse primer Taq polymerase H20_ Total comparison result The first picture is a PCR reaction solution containing BSA of the present invention Electrophoresis of PCR products after DNA amplification. The second figure is an electrophoresis diagram of a PCR product after the DNA is amplified using a conventional PCR reaction solution without BSA. Comparing the results of the first graph and the second graph, it can be found that the samples with the numbers 6 and 7 in the first graph have obvious bright bands, and the second graph has no obvious bright bands at all. This result shows that the amount of the PCR product of Example 1 is higher than the yield of Example 2. In other words, the addition of bsA to the composition of the PCR reaction solution can obtain a higher PCR yield, enabling low-level or difficult extraction of ΝΑΑ samples. Effectively amplified by PCR for further testing. Example 2 The DNA extracted in Example 1 was diluted 10-fold. Add the DNA of the total flora in the soil sample diluted 10 times and the PCR reaction solution in Table 1 to the PCR tube. Set the PCR reaction solution, place the PCR tube in the reaction 200535244 device for reaction, and then take 10 // 1 of the PCR product for electrophoresis analysis. The result is shown in the third figure. The PCR reaction conditions and electrophoresis analysis method were the same as those in Example 1. Comparative Example 2 The DNA extracted in Example 1 was diluted 10-fold. Into the PCR tube, add the DNA of the total flora in the soil sample diluted 10 times and the PCR reaction solution in Table 2. Set the PCR reaction solution, put the pCR tube into the reactor for reaction, and then take 10 / zl of the PCR product for electrophoresis analysis. The result is shown in the third figure. The PCR reaction conditions and electrophoretic analysis method were the same as those in Example 1. Comparison result The third figure is an electrophoresis chart of a PCR product after a 10-fold dilution of DNA was amplified using the pCR reaction solution containing BSA of the present invention. The fourth picture is the electrophoresis picture of the PCR product after a 10-fold dilution using a conventional PCR reaction solution without BSA. Comparing the results of the third graph and the fourth graph, it can be found that the samples of the third graph with numbers 1, 2, 3, 4, 6, 7, 8, and 9 have obvious bright bands, and the second graph only has 2, 3, and 6 And the sample of 7 had obvious bright bands. This result shows that the amount of the PCR product of Example 3 is higher than that of the example. In other words, the addition of BSA to the composition of the PCR reaction solution can obtain a higher PCR yield, so that DNA samples with low amounts or difficult extraction can be effectively used. Further detection was performed by PCR amplification. Other Implementation Aspects All the features disclosed in this specification can be combined with other methods in any form, so that the features disclosed in the book of M can be replaced by features with the same, equivalent, or similar purpose. Therefore, all features disclosed in this specification except & explicitly emphasized are just one embodiment of many equal or similar features. According to the contents disclosed in this specification, anyone familiar with the technical field can easily and clearly understand the important features of the present invention. Without departing from the spirit and scope of the present invention, different changes and modifications can be made to the present invention to make it consistent with Different uses and situations. Therefore, other implementation aspects are also included in the scope of the following patent applications. [Schematic description] The first picture is an electrophoresis picture of the PCR product of an undiluted soil DNA sample. The PCR reaction solution is a reaction solution containing bsa according to the present invention. The second picture is the electrophoresis picture of the PCR product of the undiluted soil DNA sample. The PCR reaction solution is a traditional reaction solution that does not contain bsa. The third figure is an electrophoresis diagram of a PCR product of a soil DNA sample diluted 10 times. The PCR reaction solution is a reaction solution containing bsa according to the present invention. The fourth picture is an electrophoresis diagram of a PCR product of a soil DNA sample diluted 10 times. The PCR reaction solution is a conventional reaction solution that does not include BSA. 13