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|a eng
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| 100 |
1 |
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|a Duan, W
|e verfasserin
|4 aut
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| 245 |
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|a RT-PCR heteroduplex analysis permits differentiation of transgene and host gene expression in a transgenic animal model
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|c 2002
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|a Text
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|2 rdacontent
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|a ohne Hilfsmittel zu benutzen
|b n
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|2 rdacarrier
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|a Date Completed 29.01.2003
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|a Date Revised 28.09.2018
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|a published: Print
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|a Citation Status MEDLINE
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|a In transgenic animal models, the conservation of DNA sequences between the transgene and the host wild-type gene can complicate the evaluation of the expression of each gene. The potential for gene silencing may complicate matters further. Here we report the use of RT-PCR heteroduplex analysis to differentiate the expression of a transgene and its homologous wild-type, even when these genes are very similar in their respective DNA sequences. We designed RT-PCR primers to amplify identically sized 243-bp fragments within the DNA binding domain of the p53 gene from both human and mouse mRNA samples. Ten samples from human p53 (273H) transgenic mice and 10 samples from wild-type controls were tested. Heteroduplex bands were formed in all transgenic samples but were absent from all wild-type samples. In addition, RT-PCR heteroduplex analysis was able in one sample to differentiate a silenced transgene from its wild-type allele, without the assistance of sequencing or labeling. In summary, the RT-PCR heteroduplex analysis is easy to use and has the ability to screen a large number of samples in a short time. The RT-PCR heteroduplex analysis is especially useful for the detection of expression when a transgene and the host homologous endogenous allele are too conserved in sequence to design species-specific RT-PCR primers
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|a Research Support, U.S. Gov't, P.H.S.
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|a Technical Report
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4 |
|a Journal Article
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1 |
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|a Ding, H
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Zhu, W G
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Srinivasan, K
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Otterson, G A
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Villalona-Calero, M A
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Ottersom, G A
|e verfasserin
|4 aut
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| 773 |
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8 |
|i Enthalten in
|t BioTechniques
|d 1993
|g 33(2002), 1 vom: 12. Juli, Seite 58, 60-2, 64 passim
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
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| 773 |
1 |
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|g volume:33
|g year:2002
|g number:1
|g day:12
|g month:07
|g pages:58, 60-2, 64 passim
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