Literature DB >> 7760323

Modification of standard proteinase K/phenol method for DNA isolation to improve yield and purity from frozen blood.

N N Ahmad1, A B Cu-Unjieng, L A Donoso.   

Abstract

Research in medical genetics may frequently involve freezing of large numbers of peripheral blood samples. This is a convenient method for storing blood for subsequent DNA isolation and analysis. An area of potential concern is the low yield of DNA from blood samples that have been frozen. Here we report a modification of the widely used standard proteinase K/phenol DNA isolation method for improving the yield and purity of DNA from frozen blood samples, by an initial trypsinisation of whole blood before cell lysis to obtain lymphocytic nuclei and subsequent DNA purification. We report an increased total yield of DNA with pretrypsinised blood as well as improved purity. These results indicate that trypsinisation of thawed whole blood helps the deproteinisation process, reducing the amount of protein associated with the nuclear pellet. This modification to improve yield and purity of DNA from frozen blood samples should be useful to laboratories performing DNA based diagnostic work or studying molecular genetic mechanisms of disease.

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Year:  1995        PMID: 7760323      PMCID: PMC1050235          DOI: 10.1136/jmg.32.2.129

Source DB:  PubMed          Journal:  J Med Genet        ISSN: 0022-2593            Impact factor:   6.318


  7 in total

1.  Repeated freezing and thawing of peripheral blood and DNA in suspension: effects on DNA yield and integrity.

Authors:  K S Ross; N E Haites; K F Kelly
Journal:  J Med Genet       Date:  1990-09       Impact factor: 6.318

2.  Purification of DNA from clotted blood.

Authors:  S Taş
Journal:  Clin Chem       Date:  1990-10       Impact factor: 8.327

3.  Influence of blood withdrawal and anticoagulant on clotting activity, hematologic data, and certain rheologic measurements.

Authors:  W H Reinhart; A Haeberli; J Stark; P W Straub
Journal:  J Lab Clin Med       Date:  1990-01

4.  Interference of heparin with the polymerase chain reaction.

Authors:  E Beutler; T Gelbart; W Kuhl
Journal:  Biotechniques       Date:  1990-08       Impact factor: 1.993

5.  Guidelines for DNA banking. Report of the Clinical Genetics Society working party on DNA banking.

Authors:  J R Yates; S Malcolm; A P Read
Journal:  J Med Genet       Date:  1989-04       Impact factor: 6.318

6.  Analysis of human Y-chromosome-specific reiterated DNA in chromosome variants.

Authors:  L M Kunkel; K D Smith; S H Boyer; D S Borgaonkar; S S Wachtel; O J Miller; W R Breg; H W Jones; J M Rary
Journal:  Proc Natl Acad Sci U S A       Date:  1977-03       Impact factor: 11.205

Review 7.  DNA isolation and Southern analysis: a clinician's view.

Authors:  M Borst; D M Miller
Journal:  Am J Med Sci       Date:  1990-05       Impact factor: 2.378

  7 in total
  5 in total

1.  Antifreeze solution improves DNA recovery by preserving the integrity of pathogen-infected blood and other tissues.

Authors:  D S Leal-Klevezas; I O Martínez-Vázquez; B Cuevas-Hernández; J P Martínez-Soriano
Journal:  Clin Diagn Lab Immunol       Date:  2000-11

2.  Lack of association between integrin αvβ3 gene polymorphisms and hemorrhagic fever with renal syndrome in Han Chinese from Hubei, China.

Authors:  Xiao-Ping Chen; Hai-Rong Xiong; Ni Zhu; Qing-Zhou Chen; Hui Wang; Chao-Jie Zhong; Mei-Rong Wang; Shuang Lu; Fan Luo; Wei Hou
Journal:  Virol Sin       Date:  2017-02-09       Impact factor: 4.327

3.  Enzymatic Antioxidant Defense and Polymorphic Changes in Male Infertility.

Authors:  Jędrzej Baszyński; Piotr Kamiński; Maria Bogdzińska; Sławomir Mroczkowski; Marek Szymański; Karolina Wasilow; Emilia Stanek; Karolina Hołderna-Bona; Sylwia Brodzka; Rafał Bilski; Halyna Tkachenko; Natalia Kurhaluk; Tomasz Stuczyński; Małgorzata Lorek; Alina Woźniak
Journal:  Antioxidants (Basel)       Date:  2022-04-22

4.  Correlation between the -1562C/T polymorphism in the matrix metalloproteinase-9 gene and hemorrhagic transformation of ischemic stroke.

Authors:  Xiaoman Zhang; Xinhui Cao; Xiaoyu Xu; Aifan Li; Yuming Xu
Journal:  Exp Ther Med       Date:  2015-01-16       Impact factor: 2.447

5.  Xmn1-158 γGVariant in B-Thalassemia Intermediate Patients in South-East of Iran.

Authors:  Ebrahim Miri-Moghaddam; Sara Bahrami; Majid Naderi; Ali Bazi; Morteza Karimipoor
Journal:  Int J Hematol Oncol Stem Cell Res       Date:  2017-04-01
  5 in total

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