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|a DE-627
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|a eng
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1 |
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|a Bercovich, J A
|e verfasserin
|4 aut
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|a Effect of DNA concentration on recombinant plasmid recovery after blunt-end ligation
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|c 1992
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|a Text
|b txt
|2 rdacontent
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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|a Band
|b nc
|2 rdacarrier
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|a Date Completed 05.08.1992
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|a Date Revised 23.11.2016
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|a published: Print
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|a Citation Status MEDLINE
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|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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|a Comparative Study
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|a Journal Article
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|a DNA, Recombinant
|2 NLM
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|a DNA Ligases
|2 NLM
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|a EC 6.5.1.-
|2 NLM
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1 |
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|a Grinstein, S
|e verfasserin
|4 aut
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700 |
1 |
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|a Zorzopulos, J
|e verfasserin
|4 aut
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773 |
0 |
8 |
|i Enthalten in
|t BioTechniques
|d 1988
|g 12(1992), 2 vom: 01. Feb., Seite 190, 192-3
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
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773 |
1 |
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|g volume:12
|g year:1992
|g number:2
|g day:01
|g month:02
|g pages:190, 192-3
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