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- W1943104007 abstract "Abstract The diagnosis of familial defective apolipoprotein B-100 (FDB) has been facilitated by the use of mutagenic polymerase chain reaction (PCR) primers to introduce restriction sites at the FDB gene locus. We describe a two-test strategy for diagnosing FDB that overcomes the potential for error in single-test methods based on such techniques. We introduce an Sau96I restriction site for PCR products of the normal apolipoprotein B allele. Incomplete digestion of the PCR product with Sau96I suggests an FDB heterozygote, although a false-positive result due to nonideal digestion conditions remains a possibility. In such cases we use a second test that introduces an ScaI restriction site in PCR products of the FDB allele. The diagnosis is confirmed if half of this product can be digested with ScaI. Both tests use 0.25 units of Taq polymerase and are robust with respect to annealing temperature (31-58 degrees C) and to Mg2+ concentration (1.0-3.2 mmol/L)." @default.
- W1943104007 created "2016-06-24" @default.
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- W1943104007 date "1993-01-01" @default.
- W1943104007 modified "2023-09-24" @default.
- W1943104007 title "A robust strategy for screening and confirmation of familial defective apolipoprotein B-100" @default.
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- W1943104007 doi "https://doi.org/10.1093/clinchem/39.1.118" @default.
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