한컴오피스 
목차
Ⅰ. PCR의 일반적인 thermal programmingⅡ. PCR기법의 응용
Ⅲ. References
본문내용
Ⅰ. PCR의 일반적인 thermal programming.1. Hot Start
PCR은 동일한 parameter로 구성된 반복 cycle들을 통해서 이루어진다. 이것은 PCR machine의 thermal parameter가 반복된다는 의미이다. PCR reaction은 매 cycle마다 달라지게 된다. PCR의 components와 pH 등은 역동적으로 변화하면서 매번 새로운 환경으로 cycle을 지난다. 크게 이 역동적인 cycle들은 initial screening phase와 exponential replication phase 두 가지로 나누어 생각할 수 있다.
1) Initial screening phase에서는 primer가 target으로 하는 sequences를 찾아서 annealing하고 원하는 size의 target fragment를 만들어 내는 phase.
2) exponential replication phase는 만들어진 target fragment를 계속 복제해내는 phase.
이렇게 두가지로 크게 나누는 이유는 initial screening phase에서 PCR의 scnsitivity 혹은 yield와 specificity가 결정이 되기 때문이다. 이 phase에서 primer mismatch나 partial binding에 의해서 잘못된 fragments가 생겨나면 이후에서는 원하는 target fragment와 동일한 조건에서 같이 증폭이 되어 버리기 때문이다.
처음 annealing step에서 온도 값이 적절하다면 primer는 정확히 자기의 matching sequences에 binding합니다. 하지만 Tm값보다 낮은 온도에서 annealing을 시킬 경우 31 end의 부분적인 sequences만 matching 되더라도 primer의 partial binding이 일어난다.
참고 자료
Junhong Min and Antje J. Baeumner1 (2002) Highly Sensitive and Specific Detection of Viable Escherichia coli in Drinking Water. Analytical Biochemistry 303, 186-193Barry Schweitzer* and Stephen Kingsmore (2001) Combining nucleic acid amplification and detection. Current Opinion in Biotechnology 2001,12:21-27
Anderson S, Bruijn MHL, Coulson AR, Eperon IC, Sanger F and Young IG (1982) Complete sequence of bovine mitochondrial DNA: Conserved features of the mammalian mitochondrial genome. J Mol Biol 156:683-717.
Attardi G (1985) Animal mitochondrial DNA: An extreme example of genetic economy. Int Rev Cytol 93:93-145.
Bell BR, McDaniel BT and Robison OW (1985) Effects of cytoplasmic inheritance on production traits of dairy cattle. J Dairy Sci 68:2038-2051.
Birch-Machin MA (2006) The role of mitochondria in ageing and carcinogenesis. Clin Exp Dermatol 31:548-552.
Birky CW (2001) The inheritance of genes in mitochondria and chloroplasts: Laws, mechanisms, and models. Annu Rev Genet 35:125-148.
Boettcher PJ, Freeman AE, Johnston SD, Smith RK, Beitz DC and McDaniel BT (1996a) Relationships between polymorphism for mitochondrial deoxyribonucleic acid and yield traits of Holstein cows. J Dairy Sci 79:647-654.
Boettcher PJ, Kuhn MT and Freeman AE (1996b) Impacts of cytoplasmic inheritance on genetic evaluations. J Dairy Sci 79:663-675.
Boldman K, Van Vleck L, Kriese L and Kachman S (1995) MTDFREML: User's Guide. Departament of Agriculture/Agricultural Research Service, Lincon.
Brandon MC, Lot MT, Nguyen KC, Spolim S, Navathe SB, Baldi P and Wallace DC (2005) MITOMAP: A human mitochondrial genome database-2004 update. Nucleic Acids Res 33:D611-613.
Garnero AV, Lôbo RB, Bezzerra LAF and Oliveira HN (2001) Comparação de alguns critérios de seleção para crescimento na raça nelore. Rev Bras Zootec 30:714-718. Abstract in English.
Gibson JP, Freeman AE and Boettcher PJ (1997) Cytoplasmic and mitochondrial inheritance of economic traits in cattle. Livest Prod Sci 47:115-124.
Gunski RJ, Bezzerra LAF, Meirelles FV and Lôbo RB (2001) Efeito do DNA mitocondrial sobre características de crescimento pré-desmama na raça Nelore. In: Anais da 38� Reunião Anual da Sociedade Brasileira de Zootecnia, Piracicaba, pp 23-26. Abstract in English.
Mannen H, Kojima T, Oyama K, Mukai F, Ishida T and Tsuji, S (1998) Effect of mitochondrial DNA variation on carcass traits of Japanese Black cattle. J Anim Sci 76:36-41.
Mannen H, Morimoto ML, Oyamat K, Mukai F and Tsuji S (2003) Identification of mitochondrial DNA substitutions related to meat quality in Japanese Black cattle. J Anim Sci 81:68-73.
Martins-Filho R and Lôbo RB (1991) Circunferência escrotal como critério de seleção em bovinos tendo em vista o melhoramento de características reprodutivas em ambos os sexos. Ciênc Animal 1:83-91. Abstract in English.
Meirelles FV, Rosa AJM, Lôbo RB, Garcia JM, Smith LC and Duarte FM (1999) Is the American Zebu really Bos indicus? Genet Mol Biol 22:543-546.
Northcutt SL, Willham RL and Wilson DE (1991) Genetic parameters for nuclear and non-nuclear inheritance in three synthetic lines of beef cattle differing in mature size. J Anim Sci 69:4745-4753.
Pegoraro L, Yang Z, Samake S, Meirelles FV, Bordingnon V, Moquin LC and Smith LC (1996) Sequence comparison of mitochondrial tRNA genes and origin of light strand replication in Bos taurus and Nelore (Bos indicus) breeds. Anim Genet 27:91-94.
Pereira E, Eler JP and Ferraz JP (2002) Análise genética de características reprodutivas na raça Nelore. Pesq Agropec Bras 37:703-708. Abstract in English.
Quintanilla R, Varona L, Pujol MR and Piedrafita J (1999) Maternal animal model with correlation between maternal environmental effects of related dams. J Anim Sci 77:2904-2917.
Ron M, Yoffe O and Weller JI (1993) Sequence variation in D-loop mtDNA of cow lineages selected for high and low maternal effects on milk production. Anim Genet 24:183-186.
Sambrook J, Fritch EF and Maniatis T (2001) Molecular Cloning: A Laboratory Manual. 3 edition. Cold Spring Harbor Laboratory Press, New York.
SAS Institute Inc. (2001) Statistical Analysis Software. 8.2 edition. SAS Institute Inc., Cary.
Schutz MM, Freeman AE, Beitz DC and Mayfield JE (1992) The importance of maternal lineage on milk yield traits of dairy cattle. J Dairy Sci 75:1331-1341.
Schutz MM, Freeman AE, Lindberg GL and Beitz DC (1993) Effects of maternal lineages grouped by mitochondrial genotypes on milk yield and composition. J Dairy Sci 76:621-629.
Schutz MM, Freeman AE, Lindberg GL, Koehler CM and Beitz DC (1994) The effect of mitochondrial DNA on milk production and health of dairy cattle. Livestock Production Science 37:283-295.
Silveira JC, McManus C, Mascioli AS, Silva LOC, Silveira AC, Garcia JAS and Louvandini H (2004) Study of genetic and environmental factors on production and reproduction traits in a Nelore herd in Mato Grosso do Sul state. Rev Bras Zootec 33:1432-1444. Smith LC and Alcivar AA (1993) Cytoplasmic inheritance and its effects on development and performance. J Reprod Fertil Suppl 48:31-43.
Smith LC, Bordignon V, Garcia JM and Meirelles FV (2000) Mitochondrial genotype segregation and effects during mammalian development: Applications to biotechnology. Theriogenology 53:35-46.
Sutarno CG, Cummins JM, Greeff J and Lymbery AJ (2002) Mitochondrial DNA polymorphisms and fertility in beef cattle. Theriogenology 57:1603-1610.
Suzuki R, Kemp SJ and Teale AJ (1993) Polymerase chain reaction analysis of mitochondrial DNA polymorphism in N'Dama and Zebu cattle. Anim Genet 24:339-343.
Taanman JW (1999) The mitochondrial genome: Structure, transcription, translation and replication. Biochim Biophys Acta 1410:103-123.
Tess MW and MacNeil MD (1994) Evaluation of cytoplasmic genetic effects in Miles City Line 1 Hereford cattle. J Anim Sci 72:851-856.
Tess MW, Reodecha C and Robison OW (1987) Cytoplasmic genetic effects on preweaning growth and milk yield in Hereford cattle. J Anim Sci 65:675-684.
Tess MW and Robison OW (1990) Evaluation of cytoplasmic genetic effects in beef cattle using an animal model. J Anim Sci 68:1899-1909.
