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231221s1992 xx ||||| 00| ||eng c |
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|a pubmed25n0054.xml
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|a (DE-627)NLM015981975
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|a (NLM)1616702
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Kaltenboeck, B
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Efficient production of single-stranded DNA as long as 2 kb for sequencing of PCR-amplified DNA
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| 264 |
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1 |
|c 1992
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| 336 |
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|a Text
|b txt
|2 rdacontent
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| 337 |
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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| 338 |
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|a Band
|b nc
|2 rdacarrier
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| 500 |
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|a Date Completed 05.08.1992
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|a Date Revised 14.11.2007
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|a published: Print
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|a Citation Status MEDLINE
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|a A modification of the asymmetric PCR method is described, which reliably facilitates sequencing of PCR-amplified DNA. This procedure produces single-stranded DNA fragments as long as two kilobases that are suitable for dideoxy DNA sequencing. First, a PCR for double-stranded DNA is preformed under optimal conditions (double-stranded PCR). Then, a 5-10-microliters fraction of the double-stranded PCR and a single primer are used to generate single-stranded DNA in a separate PCR (single-stranded PCR). The concentration of the single primer are used to generate single-stranded DNA in a separate PCR (single-stranded PCR). The concentration of the single primer is approximately 0.4 microM. Usually 15 to 25 cycles of single-stranded PCR are optimal to produce single-stranded DNA for four to eight sequencing reactions. The single-stranded DNA is purified by centrifugal ultrafiltration and used directly in dideoxy sequencing. This method was employed to produce high-quality single-stranded DNA templates from a variety of organisms for efficient DNA sequencing of PCR-amplified DNA
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| 650 |
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|a Journal Article
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| 650 |
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4 |
|a Research Support, Non-U.S. Gov't
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| 650 |
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4 |
|a Research Support, U.S. Gov't, Non-P.H.S.
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| 650 |
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4 |
|a Research Support, U.S. Gov't, P.H.S.
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| 650 |
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7 |
|a DNA, Single-Stranded
|2 NLM
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| 650 |
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|a DNA
|2 NLM
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| 650 |
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7 |
|a 9007-49-2
|2 NLM
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| 700 |
1 |
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|a Spatafora, J W
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Zhang, X
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Kousoulas, K G
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Blackwell, M
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Storz, 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 164, 166, 168-71
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
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| 773 |
1 |
8 |
|g volume:12
|g year:1992
|g number:2
|g day:01
|g month:02
|g pages:164, 166, 168-71
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|d 12
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|h 164, 166, 168-71
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