Human genomic DNA

The otherwise impossible task of analyzing the 4 to 8 x 109 base pairs of human genomic DNA can be simplified. Many of the 23 human chromosomes can be physically separated from each other, their DNA... 

Rudbeck L, Dissing J. Rapid, simple alkaline extraction of human genomic DNA from whole blood, buccal epithelial cells, semen and forensic stains for PCR. BioTechniques 1998 25 588-592. 

Cl. Cariello, N. F., Scott, J. K., et al, Resolution of a missense mutant in human genomic DNA by denaturing gradient gel electrophoresis and direct sequencing using in vitro DNA amphfication HPRT Munich. Am. J. Hum. Genet. 42(5), 726-734 (1988). 

Less than one-third of human genomic DNA consists of genes. Much of the remainder consists of repeated sequences of various types. Nucleic acid parasites known as transposons account for about half of the human genome. 

FIGURE 9.5 Sex determination from human genomic DNA using the PCR-CE microdevice. Top amplification from female DNA. Only the 157 bp peak representing amplification from the X chromosome is seen. Middle amplification from male DNA both the 157 bp X-chromosome and the 200 bp Y chromosome amplicons are observed. Bottom DNA sizing ladder for amplicon size reference, run separately on the same device [282]. Reprinted with permission from the Royal Society of Chemistry. 

Restriction Analysis of human genomic DNA has revealed that there are many differences... 

FGENESH 1.0 Prediction of potential genes in Human genomic DNA Seq name hmLyz... 

Itoh H, Toyama RI, Kozasa TI, Tsukamoto H, Matsuoka M, Kaziro Y. 1988. Presence of three distinct molecular spedes of G protein a subunit structure of rat cDNA and human genomic DNAs. J Biol Chem 263 6656-6664. 

Burge, C. Karlin, S. (1997). Prediction of complete gene structures in human genomic DNA. J Mol Biol 268,78-94. 

Raisi, F., Blizard, B.A., Shabari, A.R., Ching, J., Kintz, G.J., Mitchell, J., Lemoff, A., Taylor M.T., Weir, F., Western, L., Wong, W., Joshi, R., Howland, P., Chauhan, A., Nguyen, P., Peterson, K.E. (2004). Human genomic DNA analysis using a semi-automated sample preparation, amplification, and electrophoresis separation platform. J. Separation Sci. 27 275-83. 

Seeger C, Batz H-G, 0rum H. PNA-mediated purification of PCR amplifiable human genomic DNA from whole blood. BioTechniques 1997 23 512-517. 

P2Y4 receptors (P2v< uridine receptors pyrimidine nucleotide receptors) have been cloned from human genomic DNA they couple to the InsPs/DAG system (which in this instance is Pertussis-toxin-sensitive). Agonist show a rank order of potency UTP > UDP > ATP (partial agonist). Little is known, as yet, about the distribution or actions of these receptors, but they are found in human placenta and lung. 

Murai T, Kazikuza A, Takumi T, Ohkubo H, Nakanishi S. 1989. Molecular cloning and sequence analysis of human genomic DNA encoding a novel membrane protein which exhibits a slowly activating potassium channel activity. Biochem. Biophys. Res. Commun. 161 176-81... 

The accuracy of real-time PCR quantification depends not only on the method chosen to analyze the curves, but also on the quality of calibrators used. Purified PCR products quantified by spectrophotometry are easily obtained. When serially diluted, these calibrators can accurately quantify the amount of target in human genomic DNA. Alternatively, purified plasmids or genomic DNA can be used as calibrators. Limiting dilution analysis to determine the amount of ampMable DNA is seldom necessary. The precision of quantitative real-time PCR depends on the copy number. When the initial target concentration is low, imprecision is high. Part of the variance comes from stochastic limitations as defined by the Poisson distribution as described earlier. In addition, the PCR efficiency may be more variable at low copy numbers. 

Burge C. (1997) Identification of genes in human genomic DNA. Ph.D. Thesis, Stanford, 152p. 

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