Chloroplast deoxyribonucleic acid Isolation: purity achieved without nuclease digestion

1981 ◽  
Vol 59 (11-12) ◽  
pp. 911-915 ◽  
Author(s):  
Patsy R. Rhodes ◽  
S. D. Kung
Keyword(s):  
Chloroplast Dna ◽  
Liquid Nitrogen ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Buoyant Density ◽  
Silica Sol ◽  
Plant Tissues ◽  
Nuclease Digestion ◽  
Density Analysis ◽  
Digestion Step

Chloroplasts, obtained from plant tissues homogenized in liquid nitrogen, were freed of nuclei on silica sol gradients. Buoyant density analysis of denatured–renatured DNA and the clarity of restriction fragment patterns demonstrate the purity of these preparations. In this manner, chloroplast DNA free of substantial nuclear DNA contamination was obtained from several plant species without the use of a deoxyribonuclease digestion step.

scholarly journals Kinetic complexity of chloroplastal deoxyribonucleic acid and mitochondrial deoxyribonucleic acid from higher plants

1969 ◽  
Vol 112 (5) ◽  
pp. 777-786 ◽  
Author(s):  
Richard Wells ◽  
Max Birnstiel
Keyword(s):  
Molecular Weight ◽  
Mitochondrial Dna ◽  
Gaussian Distribution ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Higher Plants ◽  
Buoyant Density ◽  
Cell Fractionation ◽  
Caesium Chloride ◽  
Cellular Dna

1. Chloroplasts and mitochondria were isolated by aqueous and non-aqueous cell-fractionation techniques. In a variety of higher plants the mitochondrial DNA bands in a caesium chloride gradient at 1·706g.cm.−3, whereas chloroplastal DNA has a buoyant density of 1·697g.cm.−3. 2. In total cellular DNA of moderate molecular weight, the chloroplastal DNA is found within the Gaussian distribution of the nuclear DNA and is not resolved as a satellite. 3. Both chloroplastal DNA and mitochondrial DNA from lettuce renature rapidly. 4. The kinetic complexity of mitochondrial DNA is > 108 daltons. 5. Chloroplastal DNA is made up from fast and slow renaturing sequences with kinetic complexities of 3×106 and 1·2×108 daltons respectively. 6. From the discrepancy between analytical and kinetic complexity it is concluded that chloroplastal DNA is extensively reiterated.

scholarly journals Mitochondrial deoxyribonucleic acid from Tetrahymena pyriformis and its kinetic complexity

1970 ◽  
Vol 116 (5) ◽  
pp. 811-817 ◽  
Author(s):  
R. A. Flavell ◽  
I. G. Jones
Keyword(s):  
Mitochondrial Dna ◽  
Melting Temperature ◽  
Density Gradient ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Buoyant Density ◽  
Tetrahymena Pyriformis ◽  
Base Pairing ◽  
Caesium Chloride ◽  
Kinetics Of

1. Mitochondrial DNA from Tetrahymena pyriformis strain T has a buoyant density (ρ) of 1.685 compared with ρ1.688 for whole cell DNA. Mitochondrial preparations from T. pyriformis strain W show an enrichment of a light satellite (ρ1.686), although this is not obtained free from nuclear DNA (ρ1.692). 2. T. pyriformis mitochondrial DNA renatures rapidly and the kinetics of this process indicate a complexity of approx. 3×107 daltons. 3. The base-pairing in the renaturation product is of a precise nature, since the ‘melting’ temperature (80.5°C) is indistinguishable from that of the native DNA (80.5°C). 4. Centrifugation of mitochondrial DNA in an alkaline caesium chloride density gradient gives two bands, implying the separation of the complementary strands.

scholarly journals Involvement of deoxyribonucleic acid polymerase β in nuclear deoxyribonucleic acid synthesis

1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Dna Polymerase Beta ◽  
Cell Nuclei ◽  
High Molecular Weight Dna ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.

scholarly journals MINOR COMPONENTS OF THE DNA OF PHYSARUM POLYCEPHALUM

1969 ◽  
Vol 40 (2) ◽  
pp. 484-496 ◽  
Author(s):  
Charles E. Holt ◽  
Elizabeth G. Gurney
Keyword(s):  
Cell Cycle ◽  
Physarum Polycephalum ◽  
Nuclear Dna ◽  
Buoyant Density ◽  
Cell Fractionation ◽  
Minor Components ◽  
Variable Level ◽  
The Mean ◽  
S Period ◽  
Dna Components

DNA metabolism in the slime mold Physarum polycephalum was studied by centrifugation in CsCl of lysates of cultures labeled with radioactive thymidine at various times in the cell cycle. During the G2 (premitotic) phase of the cell cycle, two components of the DNA are labeled. One component is lighter (buoyant density 1.686 g/cc) than the mean of the principal DNA (1.700 g/cc), and one is heavier (approximately 1.706 g/cc). The labeled light DNA was identified chemically by its denaturability, its susceptibility to DNase, and the recovery of its radioactivity in thymine. Cell fractionation studies showed that the heavy and the principal DNA components are located in the nucleus and that the light DNA is in the cytoplasm. The light DNA comprises approximately 10% of the DNA. About ⅓–½ of the light DNA is synthesized during the S period, and the remainder is synthesized throughout G2 (there is no G1 in Physarum). The light DNA is metabolically stable. A low, variable level of incorporation of radioactive thymidine into the principal, nuclear DNA component was observed during G2.

Is There An Expiration Date For Samples Stored In Liquid Nitrogen? Lessons Learnt From Plant Tissues

Cryobiology ◽  
2021 ◽  
Vol 103 ◽  
pp. 161-162
Author(s):  
Daniel Ballesteros ◽  
Christina Walters ◽  
Valerie C. Pence ◽  
Victoria Garcia-Sakai ◽  
Hugh W. Pritchard
Keyword(s):  
Liquid Nitrogen ◽  
Plant Tissues ◽  
Expiration Date ◽  
Lessons Learnt

scholarly journals NUCLEAR DNA AND CYTOPLASMIC DNA FROM TISSUES OF HIGHER PLANTS

1965 ◽  
Vol 27 (3) ◽  
pp. 451-457 ◽  
Author(s):  
Yasuo Hotta ◽  
Alix Bassel ◽  
Herbert Stern
Keyword(s):  
Inorganic Phosphate ◽  
Nuclear Dna ◽  
Higher Plants ◽  
Soluble Fraction ◽  
Buoyant Density ◽  
Wheat Roots ◽  
Cytoplasmic Dna ◽  
Sucrose Medium

Young wheat roots were labeled with 32P-inorganic phosphate. Following the labeling period, roots were homogenized in a sucrose medium and fractionated into nuclei, cytoplasmic particles (including proplastids and mitochondria), and a soluble fraction containing most of the microsomes. DNA prepared from the particles had a higher buoyant density than that from the nuclei and showed a marked loss in total label if the roots were exposed to non-radioactive medium for 48 hours prior to fractionation of the cells.

scholarly journals The diagnostic significance of nuclear deoxyribonucleic acid measurement in automated cytology.

1979 ◽  
Vol 27 (1) ◽  
pp. 520-521 ◽  
Author(s):  
E Sprenger ◽  
S Witte
Keyword(s):  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Flow Cytometric Analysis ◽  
Nuclear Dna Content ◽  
Malignant Cell ◽  
Cell Populations ◽  
Diagnostic Significance ◽  
Flow Cytometric ◽  
Screening Systems ◽  
Nuclear Deoxyribonucleic Acid

Flow cytometric analysis of cytologic samples from four different organs shows that nuclear DNA content of malignant cell populations depends to a large extent on organ of origin of the tumor. This fact must be considered in planning screening systems.

Determination of guanine plus cytosine content in Streptococcus pyogenes

1975 ◽  
Vol 21 (5) ◽  
pp. 722-724 ◽  
Author(s):  
James G. Stuart ◽  
Joseph J. Ferretti
Keyword(s):  
Streptococcus Pyogenes ◽  
Thermal Denaturation ◽  
Buoyant Density ◽  
Density Analysis

The guanine and cytosine content of Streptococcus pyogenes DNA was determined by thermal denaturation and buoyant density analysis to be 36.7% and 38.7%, respectively.

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