Purification of human genomic DNA from whole blood using sodium perchlorate in place of phenol

Johns, Malcolm B.; Paulus-Thomas, J.

doi:10.1016/0003-2697(89)90430-2

Cited by 200 publications

(83 citation statements)

References 9 publications

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“…Amplification of the Genomic DNA-The total genomic DNA was isolated from cone venom glands using a NaClO 4 extraction procedure followed by RNase treatment (16). Primer 4 with an XbaI restriction site (as underlined in 5Ј-CGTCTAGA TGC CGC GCT GAA GGA-3Ј), corresponding to amino acid residues 1-5 (Cys-Arg-Ala-Glu-Gly), and primer 2 were used as a pair to amplify the genomic DNA of -BtX.…”

Section: Methodsmentioning

confidence: 99%

A Novel Conotoxin from Conus betulinus, κ-BtX, Unique in Cysteine Pattern and in Function as a Specific BK Channel Modulator

Fan

Chen

Zhang

et al. 2003

Journal of Biological Chemistry

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A novel conotoxin, -conotoxin (-BtX), has been purified and characterized from the venom of a wormhunting cone snail, Conus betulinus. The toxin, with four disulfide bonds, shares no sequence homology with any other conotoxins. Based on a partial amino acid sequence, its cDNA was cloned and sequenced. The deduced sequence consists of a 26-residue putative signal peptide, a 31-residue mature toxin, and a 13-residue extra peptide at the C terminus. The extra peptide is cleaved off by proteinase post-processing. All three Glu residues are ␥-carboxylated, one of the two Pro residues is hydroxylated at position 27, and its C-terminal residue is Pro-amidated. The monoisotopic mass of the toxin is 3569.0 Da. Electrophysiological experiments show that: 1) among voltage-gated channels, -BtX is a specific modulator of K ؉ channels; 2) among the K channels, -BtX specifically up-modulates the Ca 2؉ -and voltage-sensitive BK channels (252 ؎ 47%); 3) its EC 50 is 0.7 nM with a single binding site (Hill ‫؍‬ 0.88); 4) the time constant of wash-out is 8.3 s; and 5) -BtX has no effect on single channel conductance, but increases the open probability of BK channels. It is concluded that -BtX is a novel specific biotoxin against BK channels.

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Section: Methodsmentioning

confidence: 99%

A Novel Conotoxin from Conus betulinus, κ-BtX, Unique in Cysteine Pattern and in Function as a Specific BK Channel Modulator

Fan

Chen

Zhang

et al. 2003

Journal of Biological Chemistry

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“…Геномную ДНК выделяли из цельной крови больных посредством экстракции фенолом хлороформом после инкубации образцов кро ви с протеиназой К в присутствии 0,1% доде цилсульфата натрия [25].…”

Section: экспериментальная частьunclassified

Association of the Polymorphisms of the Tcf7l2 Genes With Type 2 Diabetes

Nikitin¹,

Потапов²,

Brovkin³

et al. 2014

Journal of Clinical Practice

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1ÔÃÁÓ Ôåäåðàëüíûé íàó÷íî-êëèíè÷åñêèé öåíòð ñïåöèàëèçèðîâàííûõ âèäîâ ìåäèöèíñêîé ïîìîùè è ìåäèöèíñêèõ òåõíîëîãèé ÔÌÁÀ Ðîññèè, ã. Ìîñêâà ÃÁÎÓ ÂÏÎ «Òþìåíñêàÿ ãîñóäàðñòâåííàÿ ìåäèöèíñêàÿ àêàäåìèÿ», ã. Òþìåíü С целью изучения ассоциации с сахарным диабетом типа 2 гена TCF7L2 проанализировано распределение частот аллелей и генотипов полиморфных маркеров rs7903146 и rs12255372 этого гена. В исследование вошли группы больных сахарным диабетом типа 2 и здоровых индивидов без признаков заболевания. Сравнительный анализ распределения частот аллелей и генотипов указывает на ассоциацию с этим заболеванием полиморфного маркера rs7903146 гена TCF7L2, в то время как для маркера rs12255372 не было обнаружено статистически значимой ассоциации. На основании полученных данных можно сделать вывод о том, что в русской популяции ген TCF7L2 ассоциирован с развитием СД типа 2.Ключевые слова: сахарный диабет типа 2, полиморфный маркер, генетическая предрасположенность. ASSOCIATION OF THE POLYMORPHISMS OF THE TCF7L2 GENES WITH TYPE 2 DIABETESNikitin A.G., Potapov V.A., Brovkin A.N., Lavrikova E.Yu., Khodyrev D.S., Shamhalova M.Sh., Smetanina S.A., Suplotova L.N., Shestakova M.V., Nosikov V.V., Averyanov A.V.To study the association with diabetes mellitus type 2 we performed analysis of the distribu tion of frequencies of alleles and genotypes two polymorphic markers of TCF7L2 gene. The study included groups of T2DM patients and unrelated controls of Russian origin. Analysis of the dis tribution of frequencies of alleles and genotypes of the polymorphic marker rs7903146 of TCF7L2 gene showed the presence of association with T2DM in Russian population, while for the marker rs12255372 was not found statistically significant associations with type 2 diabetes. We can con clude that in Russian population TCF7L2 gene is associated with development of T2DM.

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“…Genomic DNA was isolated from frozen peripheral blood samples by using the sodium perchlorate/chloroform extraction method ( Johns and Paulus-Thomas, 1989) and used as a template in polymerase chain reaction (PCR)-based genotyping analysis.…”

Section: Dna Isolation and Genotyping Proceduresmentioning

confidence: 99%

Assessment of Individual Susceptibility to Baseline DNA and Cytogenetic Damage in a Healthy Turkish Population: Evaluation with Lifestyle Factors

Kadıoğlu

Kocabaş

Demircigil

et al. 2012

Genetic Testing and Molecular Biomarkers

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Background: Cytogenetic biomarkers are most frequently used well-established endpoints in human population studies with their sensitivity for measuring exposure to genotoxic agents. They have an important role as early predictors of cancer risk. Identification of individual genotypes of metabolic gene polymorphisms helps to understand the modulation of cancer susceptibility by environmental exposures, such as cigarette smoking and other lifestyle factors. Aim: To evaluate individual susceptibility to chemicals, we determined individual DNA damage related to glutathione S-transferase (GST) genotypes (GSTM1, GSTT1, and GSTP1) in a Turkish population. Methods: Peripheral blood lymphocytes (PBL) and DNA samples of 127 subjects were analyzed for the presence of DNA damage, using single-cell gel electrophoresis (the Comet assay), and for cytogenetic parameters (chromosomal aberrations [CAs], bleomycin-induced CA, and a cytokinesis-blocked micronucleus assay), and the polymerase chain reaction/restriction fragment length polymorphism method, respectively. Results: Individuals carrying a GSTT1-null allele showed higher frequencies of CA and micronucleus (MN) ( p = 0.026, p = 0.003, respectively), whereas the GSTM1-null and GSTP1 mutant genotypes did not show any differences in cytogenetic parameters. Our findings demonstrated that none of the lifestyle factors (smoking, alcohol drinking, dietary habits, vitamin intake, and physical activity), except for vitamin intake ( p = 0.002), were significantly associated with the studied cytogenetic parameters. Conclusion: Our results suggest that the GSTT1 gene polymorphism may influence the baseline cytogenetic frequency in a healthy population. IntroductionT he role of genetic factors (early predictors) in determining human susceptibility to the carcinogenic effects of chemical agents has become a major research area in molecular epidemiology (Au, 2007). Biomarkers of early biologic effect include cytogenetic assays, such as chromosomal aberrations (CAs), micronucleus (MN), and the comet assay, which are the most frequently used endpoints in human population studies. Their sensitivity for measuring exposure to genotoxic agents and their role as early predictors of cancer risk have contributed to this success (Bonassi et al., 2005). CAs and cytokinesis-blocked micronucleus assay (CBMN) have value as biological dosimeters, and more recently, as predictors of cancer risk in epidemiological studies (Bonassi et al., 2004(Bonassi et al., , 2011. Mechanistic evidence linking CAs to early stages of cancer has been supported by the results of cohort studies that associated the CA frequency in peripheral lymphocytes of healthy individuals to future cancer incidence (Bonassi et al., 2008). Furthermore, it has been found that an increased MN frequency in peripheral blood lymphocytes predicts the risk of cancer in humans (Bonassi et al., 2007). The alkaline comet assay is also one of the most promising biomarkers to detect single-strand breaks (SSBs) as well as alkaline-labile sites under ...

show abstract

Purification of human genomic DNA from whole blood using sodium perchlorate in place of phenol

Cited by 200 publications

References 9 publications

A Novel Conotoxin from Conus betulinus, κ-BtX, Unique in Cysteine Pattern and in Function as a Specific BK Channel Modulator

A Novel Conotoxin from Conus betulinus, κ-BtX, Unique in Cysteine Pattern and in Function as a Specific BK Channel Modulator

Association of the Polymorphisms of the Tcf7l2 Genes With Type 2 Diabetes

Assessment of Individual Susceptibility to Baseline DNA and Cytogenetic Damage in a Healthy Turkish Population: Evaluation with Lifestyle Factors

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