Effect of DNA concentration on recombinant plasmid recovery after blunt-end ligation

We describe conditions for optimal recovery of recombinant plasmids after blunt-end ligation. It was found that one of the most critical parameters of the blunt-end ligation reaction is total DNA concentration (vector plus incoming DNA). This concentration was optimal in the range of 1-5 micrograms/...

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Veröffentlicht in:BioTechniques. - 1988. - 12(1992), 2 vom: 01. Feb., Seite 190, 192-3
1. Verfasser: Bercovich, J A (VerfasserIn)
Weitere Verfasser: Grinstein, S, Zorzopulos, J
Format: Aufsatz
Sprache:English
Veröffentlicht: 1992
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Comparative Study Journal Article DNA, Recombinant DNA Ligases EC 6.5.1.-
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245 1 0 |a Effect of DNA concentration on recombinant plasmid recovery after blunt-end ligation 
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520 |a We describe conditions for optimal recovery of recombinant plasmids after blunt-end ligation. It was found that one of the most critical parameters of the blunt-end ligation reaction is total DNA concentration (vector plus incoming DNA). This concentration was optimal in the range of 1-5 micrograms/ml of reaction mixture. Concentrations larger than 10 micrograms/ml result in strong inhibition. The optimal molar relationship between incoming DNA and vector was found to be 1 or less. Under these conditions, using dephosphorylated vector, recombinants are generated at a frequency of 10(6) colonies per microgram of insert, provided that transforming efficiency is about 5 x 10(7) colonies per microgram of plasmid DNA 
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650 4 |a Journal Article 
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700 1 |a Zorzopulos, J  |e verfasserin  |4 aut 
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