PCR技术原理

General Guidelines for Long-PCR Conditions and Enzyme MixturesEfficient Long-PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading polymerase (3' to 5' exo) is present at a lower concentration. Following the ...

Edit the program or pick up a program (No 10) if the program has been set. Reaction time for RAPD PCR. 1 cycle : Denature 95 C for 2 min 45 cycle of Denature 94 C 15 sec Anneal 40 C 30 sec Extend 72 C 90 sec 1 cycle of 72 C for 5 min 4 C hold Inoculate a touched-bacterial colony with a sterile toothpick into 100 µl of sterile H2O and voltex the tube. Boil ...

ABI7000 use notes ，131k，word文档http://www.wi.mit.edu/CMT/protocols/ABI7000RT_vCMT1.docPrimer Express ,96页，PDF格式http://www.wi.mit.edu/CMT/protocols/ABI7000RT_vCMT1.docSYBR® Green PCR Master Mix and RT-PCR ，56页，PDF格式http://www.wi.mit.edu/CMT/pr ...

SDS-PAGE: gel electrophoresis of proteinsTECHNIQUE is to set up gel plates before you mix the gel mixes. Use the THIN spacers and choose a comb--number of wells varies. Make both resolving gel and stack (NO APS or TEMED)--then add APS and TEMED to resolving gel, mix and pour about 3-3.5ml per gel. Before it polym ...

TAIL PCR反应体系和程序TAIL-PCR反应体系(20μl体系)Templet 1μl10x buffter 2.0μlMg2+ 1.5μldNTP 0.2μl (10mmol)P1 0.2μl (15μmol)P2(AD) 1.3μl (30μmol)Taq 0.2μl (5U/μl)ddH2O 15.1μlTail-PCR反应程序及条件Program used for Tail-PCR amplification第一次扩增1st amplification第二次扩增2ed amplif ...

幽门螺杆菌的基因诊断　　早在19世纪未，就有注意到胃粘膜组织中存在一种螺旋形微生物，其后陆续有类似的报道，但均未引起人们的足够重视.1983年，澳大利亚学者Wanen和Marshall报告，在人胃粘膜活检组织中分离出这种细菌，即幽门螺杆菌(HelicobacterPYlor;HP;原名幽门变曲菌CampylobacterPylor;CP)，并认为该菌可能是慢性胃炎和消化性溃疡的病原菌.随后各国学者相继进行研究，欧洲九 ...

个体配子DNA序列的PCR分析 前言 　　高等生物遗传图谱的构建依赖于选择性杂交后代的分析或者通过家系分析法来计 算连锁关系。对人类而言仅后者是可行的。使用长度多态性限制片段（RFLPS）在构 建人连锁图谱方面已取得长足的进步。为了对带有与已知表现型相关的RFLP标记的基 因进行定位，首先得建立间隔约10CM的遗传标记束（平均1CM等于1％重组），这样没 有基因离标记RFLP的距离会超过5CM。可靠的统计表明家系分析能够检测的遗传间距 精 ...

AluPCR:用重复序列引物 扩增来源复杂的人DNA 引言 　　聚合酶链反应PCR使不同来源的特异核酸片段的分离及分析发生了革命，但应用PCR分 离，分析特定DNA区域需要了解靶区域的边侧序列，这使扩增局限于已知DNA序列。我 们研究了从复杂的人和其他种DNA混合物中扩增未知DNA序列。特别是，我们应用PCR 分离出了在啮齿类细胞背景中含有人染色体片段的体细胞杂合体中的人DNA。使存留 在杂合子中的人特异区域序列得到分离和鉴定，避免了克 ...

PCR在结核杆菌诊断中的应用 　　结核杆菌可以导致全身性疾病，特别是在发展中国家，其发病率较高，危害性极大.近几年结核杆菌的变异性较大，对抗痨药物的耐药性增加，从而使结核病的发病大幅度增高.虽然传统的涂片抗酸染色、细菌培养以及胸片检查对大部分结核能作出正确的诊断，但对某些病人亦可造成误诊或漏诊;动物接种虽特异性较高，但因时间较长，不能满足临床快速诊断的需要.近几年也出现了几种快速诊断方法，但也存在较大的缺陷，如短期培养 ...

生物芯片在线 上一篇：聚合酶链反应在HIV感染研究中的应用 下一篇：PCR技术在DNA工程中的应用

RAPD PCR Colony Miniprep Edit the program or pick up a program (No 10) if the program has been set. Reaction time for RAPD PCR. 1 cycle : Denature 95 C for 2 min 45 cycle of Denature 94 C 15 sec Anneal 40 C 30 sec Extend 72 C 90 sec 1 cycle of 72 C for 5 min 4 C hold Inoculate a touched-bacterial colony with a sterile toothpick into 100 µl of sterile H ...

BEST" PCR conditions for amplifying DNA from plasmids25 ng linear template (~6.5 kb)50 pmol each primer100 pmol each dNTP1X Promega Taq buffer (no Mg++)1.5 mM MgCl21 U Taq DNA polymerase in 50 ul final 92°C / 2' 92°C / 30" 50°C or 55°C (depends on Tm of oligos) /30" 72°C / about 2' per kb Go to 2, 15 times 70°C/ 8' 4°C...hold. Takes about 2 ho ...

Polymerase Chain Reaction (PCR)(adapted from Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma 73019 broe@ou.edu)The amplification of DNA fragments using the polymerase chain reaction (33) is performed in either the Perkin-Elmer ...

Cepko/Tabin Lab, Harvard University RT-PCR : semi-quantitative http://axon.med.harvard.edu/~cepko/protocol/mike/R4.html 上一篇：BEST" PCR 下一篇：M. hyopneumoniae PCR

1. Because of the high sensitivity of this protocol, it is recommended that the outer PCR run in a single tube format and followed by either single tube or strip format for inner PCR. This is to reduce the possible cross contamination from one tube to the other.2. The following solutions need to be prepared:

Expand High Fidelity PCR (Boehringer)Steve Hahn Last modified 12/12/99Expand is a mixture of Taq and Pwo DNA polymerases (Boehringer). Pwo polymerase has 3' to 5' exo proofreading activity. If amplifying a 1 Kb fragment for 20 cycles, expect ~8% of the products will have at least one mutation. See Boeh ...

Protocol for Enhancing PCR of Very Difficult RegionsZiyun Yao, Shaoping Lin, HongMin Wu and B.A. Roe02-14-2002Target DNA (~5 ng/ul) 3ulGeneAmp 10x PCR buffeundefined 10ul25 mM MgCl2 (Applied Biosystems 58002032-01) 10ul7-deaza dGTP (Roche part No.92284126), dATP, dCTP, dTTP miundefined_10ulPrimer_pair__...

George Church Lab, Harvard Medical SchoolPCR_protocol.html"http://twod.med.harvard.edu/labgc/estep/longPCR_protocol.htmlEfficient Long PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofre ...

Yeast Colony PCR(Hahn lab) 9/17/02This method is more reliable than the old method of adding yeast directly to PCR tubes. No more than 1 microliter of the crude DNA should be added to the 50 microliter PCR reaction because the SDS in the DNA prep can inhibit the PCR reaction. Note: These elongation times and ann ...

Kitto Lab, The University of Texas at Austin PCR.html"http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Microbiology/PCR.htmlThis procedure is a "hot start" PCR cycle for use with large primers. PCR mass-produces DNA contained in the source plasmid between the ...