Name: polymerase chain reaction: basic protocol plus troubleshooting and optimization strategies
Uploaded: 2012-05-22T12:00:00.000+00:00
Duration: 9 min
Description: Watch this Scientific Journal Video about Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies at JoVE.com

試薬を設定し、ゲルを注ぐとカリフォルニア大学ロサンゼルス校（UCLA）エリン·サンダースにインスピレーション、ガイダンス、およびサポートのために、原稿を校正するためのカリフォルニア大学ロサンゼルス校（UCLA）クリスReddiに感謝します。私もGAL3遺伝子を増幅するために酵母ゲノムDNAとプライマーを供給するためのジャンカルロCostagutaとグレゴリー·S.ペインに感謝したいと思います。 4 - 私もラボ機器および図2を作るために使用する試薬の写真を撮るためにBhairav​​シャーに感謝します。このプロジェクトの資金は、HHMI（ハワードグラント番号52006944）によって提供されていました。

<ol>
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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formamide+can+dramatically+improve+the+specificity+of+PCR.">Formamide can dramatically improve the specificity of PCR.</a> Nucleic Acids Res. 18, 7465 (1990).</li><li>Smit, V. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=KRAS+codon+12+mutations+occur+very+frequently+in+pancreatic+adenocarcinomas.">KRAS codon 12 mutations occur very frequently in pancreatic adenocarcinomas.</a> Nucleic Acids Res. 16, 7773-7782 (1988).</li><li>Wilson, I. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+and+facilitation+of+nucleic+acid+amplification.">Inhibition and facilitation of nucleic acid amplification.</a> Appl. Environ. 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私は、開示することは何もありません。

PCRは、生物科学兵器で欠かせないツールとなっています。 PCRは、さらに生化学的な分析のためのゲノムと多様性だけでなく、単にクローニング遺伝子の洞察を得て、生物学者はゲノムを支配する力を得て、具体的かつ正確な臨床試験を可能にする、新たな機能を備えたハイブリッド遺伝子を加えることができ、科学のコースを変更した。 PCRアプリケーションは、その力を行使する科学者の想像力によって制限されます。そこにPCRの新しい特殊な使用法を説明し、多くの書籍や論文があり、多くは生物科学の次の世代にわたって開発されます。しかし、関係なく、予想されるアプローチは、基本的なフレームワークが変わっていません。 PCRは、そのすべての壮大さには、in vitroでのアプリケーション内の DNAの特定のセグメントを大量に生成することです。 PCR実験を設計する思想と忍耐を必要とします。図3に示す結果は、主要なチャンネルの一つを例示PCRの最適化戦略を設計する際にallenges。 PCRの1つのパラメータが変更されているつまり、それは相互に影響を与える可能性があります。例として、初期のPCRは最適なMgを2でサブ最適なアニーリング温度（58℃）で実施された場合、2.0 mMの+濃度、その結果は図3cに見られるように塗抹標本を生成します。塗抹標本を解決しようとすると、2.0 mMのMgCl 2を含む反応とPCR条件を設定し、61℃にアニール温度を調整伴うかもしれないしかし、図3aに示すように、これは任意の生成物を得ませんでした。したがって、むしろ誤った結果のトラブルシューティングを行うときに、単一の反応に1つの濃度を追加することなく、試薬を滴定することをお勧めします。また、PCR実験を最適化するために必要とされる最も一般的な調整は、Mg 2 +濃度を変更し、アニール温度を修正することです。ただし、これらの変更は最小限に抑えたり、異常な結果を廃止、additiの滴定しない場合VESおよび/または表2-6に説明したようにサイクリング条件のプロトコルを変更するには、問題を軽減することがあります。他のすべてが失敗した場合は、プライマーを再設計して、もう一度試してみてください。

<table border="1"><tbody><tr><td> 試薬の名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td> Taqポリメラーゼ Polymeras </td><td>シグマ</td><td> D4545-25UN </td><td></td></tr><tr><td>のdNTP </td><td>キアゲン</td><td> 201225 </td><td></td></tr><tr><td> 0.2ミリリットル薄壁チューブPCRチューブ</td><td>バイオラッド社</td><td> 223-9469 </td><td></td></tr><tr><td> 0.2ミリリットル薄壁管のキャップ</td><td>バイオラッド社</td><td> 223-9472 </td><td></td></tr><tr><td> TEバッファー： </td><td></td><td></td><td></td></tr><tr><td> EDTA二ナトリウム塩二水和物</td><td>シグマ</td><td> E5134 </td><td></td></tr><tr><td> Trizma-HClを</td><td>シグマ</td><td> T-3253 </td><td></td></tr><tr><td>カスタムファージ入門</td><td>インビトロジェン</td><td colspan="2"> 25441F_RL6_Contig2_68k TACTGCAACGCGATGTTGCG </td></tr><tr><td>カスタムファージ入門</td><td>インビトロジェン</td><td colspan="2"> 26007R_RL6_ Contig2_68k TCACCATCAATCGTGGCGGT </td></tr><tr><td>カスタム酵母入門</td><td>統合されたDNAの技術</td><td colspan="2">のSacI-Gal3（-256）ACCTGGAGCTCCTATTT TAACATGTGGATTTCTTGAAAGAATGAAATCG </td></tr><tr><td>カスタム酵母入門</td><td>統合されたDNAの技術</td><td colspan="2"> Gal3（1848）をSacI CGGATGGAGCTCGCGT GAAGGTAATTTTTTTTTATGTCTCCG </td></tr><tr><td>サーマルサイクラー</td><td>バイオラッド社</td><td> 170-9703 </td><td>私のサーマルサイクラー</td></tr><tr><td>アガロース</td><td> ISC BioExpress </td><td> E-3120-500 </td><td></td></tr><tr><td>ゲル電気泳動</td><td> Hoeffer科学機器</td><td>モデルHE33 </td><td></td></tr><tr><td> 1 kbのDNAラダー</td><td>ニューイングランドバイオラボ</td><td> N3232S </td><td></td></tr><tr><td>バイオラッド社パワーパック</td><td>バイオラッド社</td><td> 164-5050 </td><td> PowerPacは基本的な</td></tr><tr><td>ゲルドクシステム</td><td> Fotodyne株式会社</td><td colspan="2"> FOTO /アナ捜査FXワークステーション</td></tr></tbody></table>

polymerase chain reaction: basic protocol plus troubleshooting and optimization strategies

生物学の発見の黄金時代に規律をカタパルト技術の進歩がありました。例えば、微生物学の分野は​​、科学者が初めての原核生物を可視化することができアントン·ファン·レーウェンフックの顕微鏡の出現で形質転換した。ポリメラーゼ連鎖反応（PCR）の開発は、その影響は生物学で無数のsubdisciplinesにまたがると分子科学のコースを変更された技術革新の一つです。理論的なプロセスは1971年にKeppeや同僚で概説された、完全なPCR手順はシータス社の1985年の間にキャリーマリスによって記述され、実験的に適用されるまで、しかし、それは別の14年であった。この手法の自動化と洗練は、細菌サーマスのアクア 、結果的に名前のTaq DNAポリメラーゼの熱安定性DNAポリメラーゼの導入により、進行した。 PCRは、ポウです。出発材料の少量（すなわち、鋳型DNAまたは標的配列）のみからのDNAの特定のセグメント（すなわち、アンプリコン）の十分な供給を生成することができますrful増幅技術。単純な、一般的にトラブルフリーながら、誤った結果を生成する反応を複雑に落とし穴があります。 PCRが失敗したとき、それはアガロースゲル上のバンドのラダーまたはスメアとして現れ、さまざまなサイズの多くの非特異的なDNAの製品につながることができます。時々ない製品は全く形成されない。突然変異は意図せずにPCR産物の不均一な集団で、その結果、増幅産物に導入されたときに、別の潜在的な問題が発生します。忍耐と慎重なトラブルシューティングが整理し、問題（s）を解決するために採用されていない限り、PCRの失敗は、イライラになることができます。このプロトコルは、PCRの基本原理を概説し、ほとんどの標的配列の増幅をもたらす方法論を提供し、反応を最適化するための戦略を提示。このPCRガイドは、stに従うことにより、udentsのことができるようになります。 <ul><li>従来のPCR実験のための反応と熱サイクル条件を設定</li><li>様々な反応成分の機能とPCR実験で彼らの全体的な効果を理解する</li><li>任意のDNAテンプレートのPCR実験の設計と最適化</li><li>失敗したPCR実験のトラブルシューティング</li></ul>

Watch this Scientific Journal Video about Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies at JoVE.com

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

PCRは、多くの分子生物学の研究室で一般的な手法として登場しました。いくつかの従来のPCRプロトコルへのクイックガイドですここで提供される。各反応はユニークな実験であるため、製品を生成するために必要な最適な条件は異なります。反応の変数を理解することは非常に所望の結果を得ることができる機会を増やすことは、トラブルシューティングの効率を向上させます。

ポリメラーゼ連鎖反応：基本的なプロトコルプラスのトラブルシューティングと最適化戦略

In the biological sciences there have been technological advances that catapult the discipline into golden ages of discovery. For example, the field of ...

polymerase-chain-reaction-basic-protocol-plus-troubleshooting

Nanyang Technological University

Research

JoVE Journal

Biology

403.2K ビュー.  University of California, Los Angeles. PCRは、多くの分子生物学の研究室で一般的な手法として登場しました。いくつかの従来のPCRプロトコルへのクイックガイドですここで提供される。各反応はユニークな実験であるため、製品を生成するために必要な最適な条件は異なります。反応の変数を理解することは非常に所望の結果を得ることができる機会を増やすことは、トラブルシューティン?...

ビデオ: Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

The traditional strategy for the introduction of chemical functionalities is the use of solid-phase synthesis by appending suitably modified phosphoramidite precursors to the nascent chain. However, the conditions used during the synthesis and the restriction to rather short sequences hamper the applicability of this methodology. On the other hand, modified nucleoside triphosphates are activated building blocks that have been employed for the mild introduction of numerous functional groups into nucleic acids, a strategy that paves the way for the use of modified nucleic acids in a wide-ranging palette of practical applications such as functional tagging and generation of ribozymes and DNAzymes. One of the major challenges resides in the intricacy of the methodology leading to the isolation and characterization of these nucleoside analogues.
	
	In this video article, we present a detailed protocol for the synthesis of these modified analogues using phosphorous(III)-based reagents. In addition, the procedure for their biochemical characterization is divulged, with a special emphasis on primer extension reactions and TdT tailing polymerization. This detailed protocol will be of use for the crafting of modified dNTPs and their further use in chemical biology.

The protocol described herein aims to explain and abridge the numerous obstacles in the way of the intricate route leading to modified nucleoside triphosphates. Consequently, this protocol facilitates both the synthesis of these activated building-blocks and their availability for practical applications.

ヌクレオシド三リン酸 - 合成からの生化学的特徴付けへ

The  traditional strategy for the introduction of chemical functionalities is the  use of solid-phase synthesis by appending suitably modified ...

化学

Adapting 3' Rapid Amplification of CDNA Ends to Map Transcripts in Cancer

Maturation of eukaryotic mRNAs involves 3&#39; end formation, which involves the addition of a poly(A) tail. In order to map the 3&#39; end of a gene, the traditional method of choice is 3&#39; rapid amplification of cDNA ends (3&#39; RACE). Protocols for 3&#39; RACE require the careful design and selection of nested primers within the 3&#39; untranslated region (3&#39; UTR) of the target gene of interest. However, with a few modifications the protocol can be used to include the entire 3&#39; UTR and sequences within the open reading frame (ORF), providing a more comprehensive picture of the relationship between the ORF and the 3&#39; UTR. This is in addition to identification of the polyadenylation signal (PAS), as well as the cleavage and polyadenylation site provided by conventional 3&#39; RACE. Expanded 3&#39; RACE can detect unusual 3&#39; UTRs, including gene fusions within the 3&#39; UTR, and the sequence information can be used to predict potential miRNA binding sites as well as AU rich destabilizing elements that may affect the stability of the transcript.

The two different 3&#39; rapid amplification of cDNA ends (3&#39; RACE) protocols described here make use of two different DNA polymerases to map sequences that include a segment of the open reading frame (ORF), the stop codon, and the entire 3&#39; UTR of a transcript using RNA obtained from different cancer cell lines.

適応 3' CDNA の急速な拡大は、がんの成績証明書をマップする終了します。

Maturation of eukaryotic mRNAs involves 3&#39; end formation, which involves the addition of a poly(A) tail. In order to map the 3&#39; end of a gene, the ...

遺伝学

Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format

The quantitative analysis of human genetic variation is crucial for understanding the molecular characteristics of serious medical conditions, such as tumors. Because digital polymerase chain reactions (PCR) enable the precise quantification of DNA copy number variants, they are becoming an essential tool for detecting rare genetic variations, such as drug-resistant mutations. It is expected that molecular diagnoses using digital PCR (dPCR) will be available in clinical practice in the near future; thus, how to efficiently conduct dPCR with human genetic material is a hot topic. Here, we introduce a method to detect Adenomatous polyposis coli (APC) somatic mosaicism using dPCR with the chip-in-a-tube format, which allows eight dPCR reactions to be simultaneously conducted. Care should be taken when filling and sealing the reaction mixture on the chips. This article demonstrates how to avoid the over- and underestimation of positive partitions. Furthermore, we present a simple procedure for collecting the dPCR product from the partitions on the chips, which can then be used to confirm the specific amplification. We hope that this methods report will help promote the dPCR with the chip-in-a-tube method in genetic research.

Digital polymerase chain reaction (PCR) is a useful tool for the high-sensitivity detection of single nucleotide variants and DNA copy number variants. Here, we demonstrate key considerations for measuring rare variants in the human genome using digital PCR with the chip-in-a-tube format.

チップ チューブ形式を使用して散発的な家族性大腸腺腫症患者の遺伝的変異のデジタル ポリメラーゼ連鎖反応分析

The quantitative analysis of human genetic variation is crucial for understanding the molecular characteristics of serious medical conditions, such as ...

がん研究

Use of Alu Element Containing Minigenes to Analyze Circular RNAs

In addition to linear mRNAs, many eukaryotic genes generate circular RNAs. Most circular RNAs are generated by joining a 5&#39; splice site with an upstream 3&#39; splice site within a pre-mRNA, a process called back-splicing. This circularization is likely aided by secondary structures in the pre-mRNA that bring the splice sites into close proximity. In human genes, Alu elements are thought to promote these secondary RNA structures, as Alu elements are abundant and exhibit base complementarities with each other when present in opposite directions in the pre-mRNA. Here, we describe the generation and analysis of large, Alu element containing reporter genes that form circular RNAs. Through optimization of cloning protocols, reporter genes with up to 20 kb insert length can be generated. Their analysis in co-transfection experiments allows the identification of regulatory factors. Thus, this method can identify RNA sequences and cellular components involved in circular RNA formation.

Use of Alu Element Containing Minigenes to Analyze Circular RNAs

We clone and analyze reporter genes generating circular RNAs. These reporter genes are larger than constructs to analyze linear splicing and contain Alu elements. To investigate the circular RNAs, the constructs are transfected into cells and resulting RNA is analyzed using RT-PCR after removal of linear RNA.

円形RNA解析に対する小遺伝子を含むAlu要素の使用

In addition to linear mRNAs, many eukaryotic genes generate circular RNAs. Most circular RNAs are generated by joining a 5&#39; splice site with an ...

生化学

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems

This protocol describes the design of a minimal DNA template and the steps for enzymatic amplification, enabling rapid prototyping of assayable proteins in less than 24 h using cell-free expression. After receiving DNA from a vendor, the gene fragment is PCR-amplified, cut, circularized, and cryo-banked. A small amount of the banked DNA is then diluted and amplified significantly (up to 106x) using isothermal rolling circle amplification (RCA). RCA can yield microgram quantities of the minimal expression template from picogram levels of starting material (mg levels if all starting synthetic fragment is used). In this work, a starting amount of 20 pg resulted in 4 &#181;g of the final product. The resulting RCA product (concatemer of the minimal template) can be added directly to a cell-free reaction with no purification steps. Due to this method being entirely PCR-based, it may enable future high-throughput screening efforts when coupled with automated liquid handling systems.

The study describes a protocol for creating large (&#181;g-mg) quantities of DNA for protein screening campaigns from synthetic gene fragments without cloning or using living cells. The minimal template is enzymatically digested and circularized and then amplified using isothermal rolling circle amplification. Cell-free expression reactions could be performed with the unpurified product.

無細胞システムを用いたタンパク質プロトタイピングのための、最小の線形テンプレートの増幅のための迅速で酵素的な方法

This protocol describes the design of a minimal DNA template and the steps for enzymatic amplification, enabling rapid prototyping of assayable proteins ...

生物工学

Rapid Characterization of Genetic Parts with Cell-Free Systems

Characterizing and cataloging genetic parts are critical to the design of useful genetic circuits. Having well-characterized parts allows for the fine-tuning of genetic circuits, such that their function results in predictable outcomes. With the growth of synthetic biology as a field, there has been an explosion of genetic circuits that have been implemented in microbes to execute functions pertaining to sensing, metabolic alteration, and cellular computing. Here, we show a rapid and cost-effective method for characterizing genetic parts. Our method utilizes cell-free lysate, prepared in-house as a medium to evaluate parts via the expression of a reporter protein. Template DNA is prepared by PCR amplification using inexpensive primers to add variant parts to the reporter gene, and the template is added to the reaction as linear DNA without cloning. Parts that can be added in this way include promoters, operators, ribosome binding sites, insulators, and terminators. This approach, combined with the incorporation of an acoustic liquid handler and 384-well plates, allows the user to carry out high-throughput evaluations of genetic parts in a single day. By comparison, cell-based screening approaches require time-consuming cloning and have longer testing times due to overnight culture and culture density normalization steps. Further, working in cell-free lysate allows the user to exact tighter control over the expression conditions through the addition of exogenous components and DNA at precise concentrations. Results obtained from cell-free screening can be used directly in applications of cell-free systems or, in some cases, as a way to predict function in whole cells.

Well-characterized genetic parts are necessary for the design of novel genetic circuits. Here we describe a cost-effective, high-throughput method for rapidly characterizing genetic parts. Our method reduces cost and time by combining cell-free lysates, linear DNA to avoid cloning, and acoustic liquid handling to increase throughput and reduce reaction volumes.

無細胞システムによる遺伝子部分の迅速な特性評価

Characterizing and cataloging genetic parts are critical to the design of useful genetic circuits. Having well-characterized parts allows for the ...

PCR: The Polymerase Chain Reaction

The polymerase chain reaction, or PCR, is a technique used to amplify DNA through thermocycling &#8211; cyles of temperature changes at fixed time intervals. Using a thermostable DNA polymerase, PCR can create numerous copies of DNA from DNA building blocks called dNTPs. There are&#160;three steps in PCR: denaturation, annealing, and elongation. Denaturation is the first step in the cycle and causes the DNA to melt by disrupting hydrogen bonds between the bases resulting in single-stranded DNA. Annealing lowers the temperature enough to allow the binding of oligonucleotide primers to the DNA template. During the elongation step DNA polymerase will synthesize new double-stranded DNA.
This video provides an introduction to the PCR procedure. The basic principles of PCR are described as well as a step-by-step procedure for setting up a generalized PCR reaction. The video shows the necessary components for a PCR reaction, includes instruction for primer design, and provides helpful hints for ensuring successful PCR reactions.

PCR: Principle, Instrumentation, and Applications

PCR法: ポリメラーゼ連鎖反応

The polymerase chain reaction, or PCR, is a technique used to amplify DNA through thermocycling &#8211; cyles of temperature changes at fixed time ...

Education

JoVE Science Education

基礎生物学

細胞および分子生物学の基本的方法

Detecting Environmental Microorganisms with the Polymerase Chain Reaction and Gel Electrophoresis

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - Arizona University 
Demonstrating Author: Bradley Schmitz
Polymerase chain reaction (PCR) is a technique used to detect microorganisms that are present in soil, water, and atmospheric environments. By amplifying specific sections of DNA, PCR can facilitate the detection and identification of target microorganisms down to the species, strain, and serovar/pathovar level. The technique can also be utilized to characterize entire communities of microorganisms in samples.
The culturing of microorganisms in the laboratory using specialized growth media is a long-established technique and remains in use for the detection of microorganisms in environmental samples. Many microbes in the natural environment, while alive, maintain low levels of metabolic activity and/or doubling times and are thus referred to as viable but non-culturable (VBNC) organisms. The use of culture-based techniques alone cannot detect these microbes and, therefore, does not provide a thorough assessment of microbial populations in samples. The use of PCR allows for the detection of culturable microbes, VBNC organisms, and those that are no longer alive or active, as the amplification of genetic sequences does not generally require the pre-enrichment of microorganisms present in environmental samples. However, PCR cannot differentiate the aforementioned states of viability and activity. When combined with one or more culture-based techniques, the viability of certain subsets of microorganisms may still be determined.

Detecting Environmental Microorganisms Using PCR

ポリメラーゼ連鎖反応およびゲル電気泳動法と環境微生物の検出

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - Arizona University
Demonstrating Author: Bradley Schmitz
Polymerase chain reaction (PCR) is ...

環境科学

環境微生物学

Gradient Polymerase Chain Reaction to Determine the Optimum Annealing Temperature

Source: Tsujimoto, H. et al., <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/63008">Indel Detection following CRISPR/Cas9 Mutagenesis using High-resolution Melt Analysis in the Mosquito Aedes aegypti.</a> J. Vis. Exp. (2021).
This video demonstrates the gradient polymerase chain reaction for optimization of the annealing temperature of primers, which sets gradient annealing temperature along wells. The effect of different annealing temperatures is determined by the yield of amplified products that helps to optimize the reaction condition.

最適なアニーリング温度を決定するためのグラジエントポリメラーゼ連鎖反応

Source: Tsujimoto, H. et al., Indel Detection following CRISPR/Cas9 Mutagenesis using High-resolution Melt Analysis in the Mosquito Aedes aegypti. J. Vis. ...

JoVE Encyclopedia of Experiments

生物学的手法

PCR技術

Microsphere Polymerase Chain Reaction to Amplify Single-Stranded DNA

Source: Lee, S. H. et al., <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/57703">A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons.</a> J. Vis. Exp. (2018).
This video describes the technique of amplifying single-stranded DNA using oligonucleotide-coated polymer microspheres in a single-tube reaction. This process yields pure single-stranded DNA, which can be further used for DNA sequencing and microarray analysis.

一本鎖DNAを増幅するためのマイクロスフェアポリメラーゼ連鎖反応

Source: Lee, S. H. et al., A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons. J. Vis. Exp. 
...

ポリメラーゼ連鎖反応：基本的なプロトコルプラスのトラブルシューティングと最適化戦略

要約

さらに動画を探す

この動画の章

ヌクレオシド三リン酸 - 合成からの生化学的特徴付けへ

適応 3' CDNA の急速な拡大は、がんの成績証明書をマップする終了します。

チップチューブ形式を使用して散発的な家族性大腸腺腫症患者の遺伝的変異のデジタルポリメラーゼ連鎖反応分析

円形RNA解析に対する小遺伝子を含むAlu要素の使用

無細胞システムを用いたタンパク質プロトタイピングのための、最小の線形テンプレートの増幅のための迅速で酵素的な方法

無細胞システムによる遺伝子部分の迅速な特性評価

PCR法: ポリメラーゼ連鎖反応

ポリメラーゼ連鎖反応およびゲル電気泳動法と環境微生物の検出

最適なアニーリング温度を決定するためのグラジエントポリメラーゼ連鎖反応

一本鎖DNAを増幅するためのマイクロスフェアポリメラーゼ連鎖反応

ポリメラーゼ連鎖反応：基本的なプロトコルプラスのトラブルシューティングと最適化戦略

要約

さらに動画を探す

この動画の章

ヌクレオシド三リン酸 - 合成からの生化学的特徴付けへ

適応 3' CDNA の急速な拡大は、がんの成績証明書をマップする終了します。

チップ チューブ形式を使用して散発的な家族性大腸腺腫症患者の遺伝的変異のデジタル ポリメラーゼ連鎖反応分析

円形RNA解析に対する小遺伝子を含むAlu要素の使用

無細胞システムを用いたタンパク質プロトタイピングのための、最小の線形テンプレートの増幅のための迅速で酵素的な方法

無細胞システムによる遺伝子部分の迅速な特性評価

PCR法: ポリメラーゼ連鎖反応

ポリメラーゼ連鎖反応およびゲル電気泳動法と環境微生物の検出

最適なアニーリング温度を決定するためのグラジエントポリメラーゼ連鎖反応

一本鎖DNAを増幅するためのマイクロスフェアポリメラーゼ連鎖反応

チップチューブ形式を使用して散発的な家族性大腸腺腫症患者の遺伝的変異のデジタルポリメラーゼ連鎖反応分析