1. An improved procedure for extraction and analysis of cellular nucleotides
- Author
-
Hugh G. Lawford, J. Tze-Fei Wong, and Ross N. Nazar
- Subjects
Biophysics ,Acetates ,Biochemistry ,law.invention ,Acetic acid ,chemistry.chemical_compound ,Adenosine Triphosphate ,law ,Freezing ,Escherichia coli ,Methods ,Fluorometry ,Nucleotide ,Perchloric acid ,Molecular Biology ,Filtration ,chemistry.chemical_classification ,Carbon Isotopes ,Reproducibility ,Chromatography ,Nucleotides ,Extraction (chemistry) ,Phosphorus Isotopes ,Cell Biology ,Freeze Drying ,chemistry ,Yield (chemistry) ,Degradation (geology) ,Chromatography, Thin Layer - Abstract
An improved procedure is described for the extraction, separation, and determination of cellular nucleotides and related compounds. Cells, harvested by warm Millipore filtration, are extracted with 1 N acetic acid. The extract is freeze-thawed, lyophilized, and redissolved in water or buffer. It is then directly applied to precoated PEI cellulose plastic plates and concentrated in situ using a simple applicator, or directly used in an enzymic assay. This provides a 25% higher yield of nucleotides than perchloric acid, with about a 10% average deviation for replicate cell samples. The processing of the extract gives a 90% recovery, with negligible degradation and a high degree of reproducibility.
- Published
- 1970
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