| dbo:abstract |
تفاعل البوليميراز المتسلسل ذو البداية الساخنة (بالإنجليزية: Hot start PCR) هو شكل من الأشكال المعدلة من تفاعل البوليميراز المتسلسل الذي يتجنب التضخيم غير المُحدد (أو العشوائي) من الحمض النووي عن طريق تعطيل عمل التاك بوليمريز في درجة حرارة أقل. (ar) Hot start PCR is a modified form of conventional polymerase chain reaction (PCR) that reduces the presence of undesired products and primer dimers due to non-specific DNA amplification at room (or colder) temperatures. Many variations and modifications of the PCR procedure have been developed in order to achieve higher yields; hot start PCR is one of them. Hot start PCR follows the same principles as the conventional PCR - in that it uses DNA polymerase to synthesise DNA from a single stranded template. However, it utilizes additional heating and separation methods, such as inactivating or inhibiting the binding of Taq polymerase and late addition of Taq polymerase, to increase product yield as well as provide a higher specificity and sensitivity. Non-specific binding and priming or formation of primer dimers are minimized by completing the reaction mix after denaturation. Some ways to complete reaction mixes at high temperatures involve modifications that block DNA polymerase activity in low temperatures, use of modified deoxyribonucleotide triphosphates (dNTPs), and the physical addition of one of the essential reagents after denaturation. Through these additional methods, hot start PCR is able to decrease the amount of non-specific amplifications which naturally occur during lower temperatures – which remains a problem for conventional PCR. These modifications work overall to ensure that specific enzymes in solution will remain inactive or are inhibited until the optimal annealing temperature is reached. Inhibiting formation of non-specific PCR products, especially in early cycles, results in a substantial increase in sensitivity of amplification by PCR. This is of utmost importance in diagnostic applications of PCR or RT-PCR. (en) 熱啟動聚合酶鏈式反應（英語：Hot start PCR）是一種改良的聚合酶鏈式反應方法，主要用來避免操作過程中產生非特異性序列的擴增。將DNA聚合酶與其他反應物隔絕，或是使用在高熱狀況下才會活化的聚合酶，避免在未達到設定溫度前就開始反應。 熱啟動PCR有幾種方式： * 蠟隔絕法：將除了聚合酶以外的反應物加入PCR管後，加滴熔融的石蠟；待石蠟凝固後再加入聚合酶。放入PCR儀開始反應升溫後，石蠟融化浮至最頂層，聚合酶才與其他反應物混合。石蠟可同時作為防止蒸發用。 * 熱啟動聚合酶（化學型）：經過化學分子修飾的聚合酶，高熱下結構改變而啟動。 * 熱啟動聚合酶（抗體）：帶有針對聚合酶的免疫抗體，高熱下使抗體分離而啟動。 * 熱啟動聚合酶（共價鍵結）：改良自抗體型，使用小分子的化合物，阻塞聚合酶作用位置，高溫下分離。 (zh) |
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تفاعل البوليميراز المتسلسل ذو البداية الساخنة (بالإنجليزية: Hot start PCR) هو شكل من الأشكال المعدلة من تفاعل البوليميراز المتسلسل الذي يتجنب التضخيم غير المُحدد (أو العشوائي) من الحمض النووي عن طريق تعطيل عمل التاك بوليمريز في درجة حرارة أقل. (ar) 熱啟動聚合酶鏈式反應（英語：Hot start PCR）是一種改良的聚合酶鏈式反應方法，主要用來避免操作過程中產生非特異性序列的擴增。將DNA聚合酶與其他反應物隔絕，或是使用在高熱狀況下才會活化的聚合酶，避免在未達到設定溫度前就開始反應。 熱啟動PCR有幾種方式： * 蠟隔絕法：將除了聚合酶以外的反應物加入PCR管後，加滴熔融的石蠟；待石蠟凝固後再加入聚合酶。放入PCR儀開始反應升溫後，石蠟融化浮至最頂層，聚合酶才與其他反應物混合。石蠟可同時作為防止蒸發用。 * 熱啟動聚合酶（化學型）：經過化學分子修飾的聚合酶，高熱下結構改變而啟動。 * 熱啟動聚合酶（抗體）：帶有針對聚合酶的免疫抗體，高熱下使抗體分離而啟動。 * 熱啟動聚合酶（共價鍵結）：改良自抗體型，使用小分子的化合物，阻塞聚合酶作用位置，高溫下分離。 (zh) Hot start PCR is a modified form of conventional polymerase chain reaction (PCR) that reduces the presence of undesired products and primer dimers due to non-specific DNA amplification at room (or colder) temperatures. Many variations and modifications of the PCR procedure have been developed in order to achieve higher yields; hot start PCR is one of them. Hot start PCR follows the same principles as the conventional PCR - in that it uses DNA polymerase to synthesise DNA from a single stranded template. However, it utilizes additional heating and separation methods, such as inactivating or inhibiting the binding of Taq polymerase and late addition of Taq polymerase, to increase product yield as well as provide a higher specificity and sensitivity. Non-specific binding and priming or format (en) |
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تفاعل البوليمراز المتسلسل ذو البداية الساخنة (ar) Hot start PCR (en) 熱啟動聚合酶鏈式反應 (zh) |
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