J., and Rutter, W J (1979) Isolation of biologically active nbonucleic acid from sources enriched in ribonuclease. Principles and Applications (Erlich, H., ed.), Stockton, New York, pp. (1989) Detection of gene expression, in The Polymerase Chain Reaction. (1989) Diagnosis of new mutation diseases using the polymerase chain reaction, in The Polymerase Cham Reaction Principles and Applications (Erlich, H., ed.), Stockton, New York, pp. (1987) An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences Application to hemophilia A. (1989) Point mutation analysis in a human gene: Rapid preparation of total RNA, PCR amplification of cDNA, and Taq sequencing by a novel method Biotechnuiques 7, 494–499 M., Urlaub, G., Mucha, J., Grunberger, D., and Chasm, L. (1989) An improved method for directly sequencing PCR amplified material using dimethyl sulphoxide. (1988) Generation of single stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus Proc Natl. (1989) Direct genomic sequencing of alleles at the retinoblastoma locus’ Applications to carrier diagnosis and genetic counselling, in Cancer Cells: Molecular Diagnostics of Human Cancer, vol 7, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 223–227. (1987) Characterization of beta-thaiassemia mutations using direct gcnomic sequencing of amplified single copy DNA.
![complementary dna sequence complementary dna sequence](https://www.reneshbedre.com/assets/posts/revcom/revcom4.png)
(1988) Characterization of c-ki-ras oncogene alleles by direct sequencing of enzymatically amplified DNA from carcinogen-induced tumors. (1987) Length mutations in human mitochondrial DNA: Direct sequencing of enzymatic ally amplified DNA.
![complementary dna sequence complementary dna sequence](https://sciencebasedmedicine.org/wp-content/uploads/2020/11/CentralDogmaMB-768x541.png)
(1985) Enzymatic amplification of β-globin gcnomic sequences and restriction site analysis for diagnosis of sickle cell anemia. (1987) Specific synthesis of DNA in vitro via a polymnerase catalyzed chain reaction. The various approaches that have been used to overcome these problems include1.The use of 5′-end-labeled DNA-sequencing primers that are complementary to regions between the PCR primers (3) 2.Gel purification of amplified DNA to remove unwanted fragments and primer (4) 3.Spin columns for the separation of leftover primers from high mol wt material (5, 6) 4.“Asymmetric” or “unbalanced” PCR priming to generate an excess of single strands during the initial amplification (7) 5.Addition of dimethylsulfoxide (DMSO) to sequencing reactions with short annealing times (8) and 6.The use of several short, high-temperature, sequencing cycles (9). The features that can distinguish PCR products as templates for sequencing include (a) contamination of the reactions by nonspecific PCR amplification products that are complementary to the sequencing primer, (b) the persistence of “leftover” PCR primers from the amplification reactions, and (c) the potential for competition between one strand of the amplified fragment and the oligonucleotide used for the sequencing. Protocols for the sequence analysis of conventional single-stranded or double-stranded DNA templates are often unsuitable for the direct sequencing of DNA fragments generated by the polymerase chain reaction (PCR) (1,2).