Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes

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Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes. / Trujillo, L E; Pupo, E; Miranda, F; Pérez, E; Gonzalez de Valdivia, Ernesto I.

In: Revista Latinoamericana de Microbiologia, Vol. 38, No. 1, 1996, p. 31-7.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Trujillo, LE, Pupo, E, Miranda, F, Pérez, E & Gonzalez de Valdivia, EI 1996, 'Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes', Revista Latinoamericana de Microbiologia, vol. 38, no. 1, pp. 31-7.

APA

Trujillo, L. E., Pupo, E., Miranda, F., Pérez, E., & Gonzalez de Valdivia, E. I. (1996). Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes. Revista Latinoamericana de Microbiologia, 38(1), 31-7.

Vancouver

Trujillo LE, Pupo E, Miranda F, Pérez E, Gonzalez de Valdivia EI. Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes. Revista Latinoamericana de Microbiologia. 1996;38(1):31-7.

Author

Trujillo, L E ; Pupo, E ; Miranda, F ; Pérez, E ; Gonzalez de Valdivia, Ernesto I. / Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes. In: Revista Latinoamericana de Microbiologia. 1996 ; Vol. 38, No. 1. pp. 31-7.

Bibtex

@article{fed5e32053a04ef5b27c45ddefa1ca50,
title = "Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes",
abstract = "We evaluated the use of two radiolabeled lambda DNA/Hpa II substrates to detect 5'-->3', 3'-->5' single and double stranded DNA dependent exonuclease and phosphatase activities found as contaminants in restriction and modifying enzyme preparations. Looking for the meaning of the radioactive assays results in a real cloning experience, we performed a cloning simulation assay using the same conditions established for the radioactive assay (enzyme units and pmols of DNA ends). As a result, we found that for degradation percentages of the radioactive DNA substrate per enzyme unit below 0.5, the false positives in the cloning stimulation assay were less than 5%. This conditions could ensure a good performance of the enzyme preparations for cloning experiments. Finally, we described the use of the radiolabeled [gamma 33P] ATP lambda Hpa II DNA substrate to detect 5'-->3' single stranded DNA dependent exonuclease and phosphatase contaminating activities in some critical steps of the purification process of the restriction enzyme Kpn I.",
keywords = "Bacteriological Techniques, Bacteriophage lambda, Cloning, Molecular, DNA Modification Methylases, DNA Restriction Enzymes, DNA, Viral, Deoxyribonuclease HpaII, Drug Contamination, Escherichia coli, Exonucleases, Phosphoric Monoester Hydrolases, Radioactive Tracers, Sensitivity and Specificity",
author = "Trujillo, {L E} and E Pupo and F Miranda and E P{\'e}rez and {Gonzalez de Valdivia}, {Ernesto I}",
year = "1996",
language = "English",
volume = "38",
pages = "31--7",
journal = "Revista Latinoamericana de Microbiologia",
issn = "0034-9771",
publisher = "Instituto de Biotecnologia UNAM - Lic Amapola",
number = "1",

}

RIS

TY - JOUR

T1 - Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes

AU - Trujillo, L E

AU - Pupo, E

AU - Miranda, F

AU - Pérez, E

AU - Gonzalez de Valdivia, Ernesto I

PY - 1996

Y1 - 1996

N2 - We evaluated the use of two radiolabeled lambda DNA/Hpa II substrates to detect 5'-->3', 3'-->5' single and double stranded DNA dependent exonuclease and phosphatase activities found as contaminants in restriction and modifying enzyme preparations. Looking for the meaning of the radioactive assays results in a real cloning experience, we performed a cloning simulation assay using the same conditions established for the radioactive assay (enzyme units and pmols of DNA ends). As a result, we found that for degradation percentages of the radioactive DNA substrate per enzyme unit below 0.5, the false positives in the cloning stimulation assay were less than 5%. This conditions could ensure a good performance of the enzyme preparations for cloning experiments. Finally, we described the use of the radiolabeled [gamma 33P] ATP lambda Hpa II DNA substrate to detect 5'-->3' single stranded DNA dependent exonuclease and phosphatase contaminating activities in some critical steps of the purification process of the restriction enzyme Kpn I.

AB - We evaluated the use of two radiolabeled lambda DNA/Hpa II substrates to detect 5'-->3', 3'-->5' single and double stranded DNA dependent exonuclease and phosphatase activities found as contaminants in restriction and modifying enzyme preparations. Looking for the meaning of the radioactive assays results in a real cloning experience, we performed a cloning simulation assay using the same conditions established for the radioactive assay (enzyme units and pmols of DNA ends). As a result, we found that for degradation percentages of the radioactive DNA substrate per enzyme unit below 0.5, the false positives in the cloning stimulation assay were less than 5%. This conditions could ensure a good performance of the enzyme preparations for cloning experiments. Finally, we described the use of the radiolabeled [gamma 33P] ATP lambda Hpa II DNA substrate to detect 5'-->3' single stranded DNA dependent exonuclease and phosphatase contaminating activities in some critical steps of the purification process of the restriction enzyme Kpn I.

KW - Bacteriological Techniques

KW - Bacteriophage lambda

KW - Cloning, Molecular

KW - DNA Modification Methylases

KW - DNA Restriction Enzymes

KW - DNA, Viral

KW - Deoxyribonuclease HpaII

KW - Drug Contamination

KW - Escherichia coli

KW - Exonucleases

KW - Phosphoric Monoester Hydrolases

KW - Radioactive Tracers

KW - Sensitivity and Specificity

M3 - Journal article

C2 - 8783903

VL - 38

SP - 31

EP - 37

JO - Revista Latinoamericana de Microbiologia

JF - Revista Latinoamericana de Microbiologia

SN - 0034-9771

IS - 1

ER -

ID: 32642136