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Molecular Biology Problem Solver 31

Molecular Biology Problem Solver 31. Sách được nhiều nhà khoa học có uy tín, nhiều kinh nghiệm trong nghiên cứu thực nghiệm trình bày những vấn đề thường hay phát sinh trong phòng thí nghiệm. Do vậy mà sách không trình bày các protocol hay quy trình như các sách khác, thay vào đó các tác giả sẽ trình bày các vấn đề nhằm giúp giúp đọc giả: Tự nâng cao khả năng chẩn đoán nguyên nhân khi gặp các vấn để về kỹ thuật, quy trình, hóa chất, thuốc thử trong quá trình thực nghiệm trong phòng thí. | Table ll.2a Priority List Researcher I Objectives High Medium Low Quantitative H Sensitivity H Fidelity M High-throughput M Reproducibility H Cost-sensitive M Long PCR product L Limited available M starting material Short template size H Gel based L Simple method H Nonradioactivity involved H Automated H Long-term project H DNA PCR L RNA PCR H Multiple samples H Multiplex H Table ll.2b Priority List Researcher 2 Objectives High Medium Low Quantitative M Sensitivity M Fidelity H High-throughput L Reproducibility H Cost-sensitive H Long PCR product L Limited available M starting material Short template size M Gel based H Simple method L Nonradioactivity involved L Automated L Long-term project L DNA PCR L RNA PCR H Multiple samples L Multiplex L mechanisms. The purpose of Researcher 2 is to amplify the cDNA and to demonstrate the size difference by separating the two forms by gel electrophoresis. The data are needed for a manuscript due in two months. You can see the differences between the priorities and needs of the two researchers. 294 Aoyagi After setting clear objectives of what your PCR reaction must accomplish check that you have the adequate resources. This includes not only budget but also head count skill level time equipment sequence information sample supply and other issues. If time is most critical then you may require a colleague s assistance or a new instrument to do the project as quickly as possible. In a similar token if the sample is difficult to obtain in abundance the choice of PCR that minimizes the sample requirement becomes more important. Selecting one PCR strategy that optimally satisfies every research need is unlikely. At this early planning stage a compromise between competing needs will likely be required. Remember that after all the planning is complete the final PCR strategy still has to evolve at the lab bench. Identify Any Weak Links in Your PCR Strategy There are many parameters that affect the outcome of a PCR reaction. Some .