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词条创建者:admin创建时间:12-09 22:27
标签: PCR 基本实验方法

摘要:Recommended Reaction Conditions:Initial Conditions:Initial denaturation at start: 92 - 97oC for 3 - 5 min. If you denature at 97oC, denature sample only; add rest of mix after reaction colls to anneal[阅读全文:]

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词条创建者:admin创建时间:12-09 22:27
标签: PCR 基本实验方法

摘要:"Hot Start" PCR: In certain circumstances one wishes to avoid mixing primers and target DNA at low temperatures in the presence of Taq polymerase: Taq pol is almost as efficient as Klenow pol at 37oC;[阅读全文:]

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词条创建者:admin创建时间:12-09 22:27
标签: PCR 实验方法

摘要:Reagent Concentrations:Primers: 0.2 - 1.0 uMNucleotides: 50 - 200 uM EACH dNTPDimethyl sulphoxide (DMSO): 0 - 10% (v/v)Taq polymerase: 0.5 - 1.0 Units/50ul rxnTarget DNA: 1 ng - 1 ug (NB: higher concn[阅读全文:]

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词条创建者:admin创建时间:12-09 22:27
标签: Purification PCR fragments for cloning 克隆片段 纯化

摘要:After an aliquot of the PCR mixture is analyzed on an agarose gel, the remainder of the reaction is concentrated by ethanol precipitation, resuspended in buffer, and subjected to a simultaneous fill-i[阅读全文:]

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Reaction volume
词条创建者:admin创建时间:12-09 22:27
标签: Reaction volume

摘要:Q: Does the PCR reaction volume (negatively) influence the outcome?A: No, especially since the introduction of the small, thin walled, 0.2 ml plastic vials fitting the 96 well metal blocks of the ther[阅读全文:]

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Single Primer ("Semi-Random") PCR
词条创建者:admin创建时间:12-09 22:27
标签: Single Primer Semi-Random PCR

摘要:DescriptionSingle primer PCR allows amplification from known to unknown regions in chromosomes, phage, plasmids, large PCR products and other sources of DNA.At sufficiently low stringency, any primer [阅读全文:]

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词条创建者:admin创建时间:12-09 22:27
标签: REAL TIME PCR WITH SYBR GREEN I 建议PROTOCOL

摘要:1、反应体系 25ulCocktail 20 ul: Primer FP 1 ul + Primer RP 1u + 2 * SYBR GREEN I MIX 12.5 + H2O 5.5 ul cDNA 5 ul2、Primer 浓度,需要摸索,从200nM到700nm等,设置不同浓度。3、每个引物浓度,从well 1到12设置4个样[阅读全文:]

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Single tube confirmation PCR protocol
词条创建者:admin创建时间:12-09 22:27
标签: Single tube confirmation PCR protoco

摘要:The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformation along with a wild-type control. A series of fi[阅读全文:]

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Site-directed Mutagenesis using PCR
词条创建者:admin创建时间:12-09 22:27
标签: Site-directed Mutagenesis PCR

摘要:From: Molecular Biology: Current Innovations and Future Trends. Eds. A.M. Griffin and H.G.Griffin. ISBN 1-898486-01-8 1995 Horizon Scientific Press, PO Box 1, Wymondham, Norfolk, U.K. In vitro site-d[阅读全文:]

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词条创建者:admin创建时间:12-09 22:27
标签: PCR 基本实验方法

摘要:Cloning PCR ProductsT-A Cloning Strategy: Taq and other polymerases seem to have a terminal transferase activity which results in the non-templated addition of a single nucleotide to the 3'-ends of PC[阅读全文:]

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PCR技术