Nucleic Acid Isolation
Nucleic Acid Isolation - Related Videos
06:49
Isolating Nucleic Acids from Yeast
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2023
One of the many advantages to using yeast as a model system is that large quantities of biomacromolecules, including nucleic acids (DNA and RNA), can be purified from the cultured cells. This video will address the steps required to carry out nucleic acid extraction. We will begin by briefly outlining the growth and harvest, and lysis of yeast cells, which are the initial steps common to the isolation of all biomacromolecules. Next, we will discuss two unique purification methods for the...
09:52
Electricity-Free, Sequential Nucleic Acid and Protein Isolation
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Cited by 4 •
2012
A tool and chemistries are described to sequentially isolate nucleic acids followed by proteins from a sample without the need for electricity. The tool consists of a sorbent held within a transfer pipette while the isolation chemistries are based on solid-phase extraction principles. The isolated macromolecules can be analyzed by immuno-based and PCR-based assays.
02:43
Nucleic Acids
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2020
Nucleic acids are the most important macromolecules for the continuity of life. They carry the cell's genetic blueprint and carry instructions for its functioning. DNA and RNA The two main types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). DNA is the genetic material in all living organisms, ranging from single-celled bacteria to multicellular mammals. It is in the nucleus of eukaryotes and in the organelles, chloroplasts, and mitochondria. In prokaryotes, the...
06:52
Kinetic Screening of Nuclease Activity using Nucleic Acid Probes
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Cited by 5 •
2019
Altered nuclease activity has been associated with different human conditions, underlying its potential as a biomarker. The modular and easy to implement screening methodology presented in this paper allows the selection of specific nucleic acid probes for harnessing nuclease activity as a biomarker of disease.
07:58
A Polyaniline-based Sensor of Nucleic Acids
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Cited by 12 •
2016
Nucleic acids are common analytes for assessing biological systems; however, bias from enzymatic manipulation can cause concern. Here a method is described for label-free detection of nucleic acids using polyaniline. This sensitive, cost-effective sensor technology can distinguish single nucleotide differences between molecules.
FAQs
Cell lysis disrupts cells or tissues so DNA or RNA becomes accessible in the sample. The following enzymatic or chemical treatments remove proteins and other contaminants that could interfere with analysis. Keeping these stages effective is important because residual material can reduce the reliability of downstream reactions, including amplification, sequencing, cloning, and gene expression measurements.
RNA protocols must limit RNase activity because RNases can degrade RNA during handling and processing. Loss or fragmentation of the RNA changes the material available for gene expression analysis and other downstream studies. Controlling RNase activity therefore helps preserve the original sample and supports more accurate interpretation of RNA-based results.
Both approaches separate nucleic acids from the surrounding sample after lysis and contaminant removal, but they use different purification principles. Precipitation collects nucleic acids from solution, whereas selective membrane binding retains them on a membrane before elution. The chosen approach determines how the material is recovered and prepared for subsequent analysis.
The amount and condition of recovered nucleic acid strongly influence downstream accuracy and reproducibility. Incomplete lysis can limit recovery, while ineffective removal of proteins or other contaminants can interfere with analysis. For RNA, uncontrolled RNase activity adds a degradation risk. Concentration and overall purification quality should therefore be considered before interpreting experimental results.
A typical workflow begins with lysis of cells, tissues, or another biological sample. Enzymatic or chemical treatment then removes proteins and other contaminants. Nucleic acids are subsequently recovered by precipitation or by binding them to a membrane, followed by elution. The isolated material can then be assessed and directed to an appropriate downstream analysis.
The target molecule determines which handling priorities matter most. DNA isolation emphasizes recovering material suitable for applications such as polymerase chain reaction, sequencing, or cloning. RNA isolation additionally requires measures that limit RNase activity and degradation, because RNA integrity affects gene expression analysis and the interpretation of results.
Purified nucleic acids provide the starting material for polymerase chain reaction, sequencing, cloning, gene expression analysis, and molecular diagnostics. In biology and biomedical research, isolation quality affects whether these methods produce accurate and reproducible results. The procedure therefore connects biological samples to measurements used to examine nucleic acid sequences, abundance, or related molecular properties.