Nucleic acid protocols: Extraction and optimization
Biomolecule extraction, such as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from a variety of starting biological materials to be used in …
Integrated DNA and RNA extraction using magnetic beads …
The main chemical constituents for the co-extraction of RNA and DNA are GTC, DTT, sodium dodecyl sulfate (SDS), and β-mercaptoethanol 12,13,15. In the 1990s, I. Casas, L established the "GuSCN-DNA/RNA" extraction method to extract viral RNA and DNA from cerebrospinal fluid, this method was the basis for most subsequent co …
7.3: DNA Extraction
Once a sample has been obtained, the DNA must be released from the nucleus, which involves the physical disruption of the cell. After the cells have broken open, a salt solution (NaCl) and a detergent solution containing the compound SDS (sodium …
A simple and rapid method for extracting bacterial DNA from intestinal
METHODS: Five methods of extracting bacterial DNA, including Tris-EDTA buffer, chelex-100, ultrapure water, 2% sodium dodecyl sulfate and 10% Triton-100 with and without sonication, were compared with the commercial fecal DNA extraction kit method, which is considered as the gold standard for DNA extraction.
A modular method for the extraction of DNA and …
Without major adjustments, this protocol is also compatible with RNA extraction, thus enabling the efficient simultaneous extraction of DNA and RNA from …
Removal of humic substances from soil DNA using aluminium sulfate
Here we report an effective protocol for the removal of humic substance from soil DNA. The protocol involves flocculation of the humic substance by excessive Al3+, then removal of superfluous Al 3+ via pH adjustment and finally release of soil microbial DNA by SDS lysis. This technique is superior to that employed by the UltraClean Soil DNA Kit ...
DNA Extraction
After DNA extraction the DNA is generally a series of large fragments averaging 25,000 to 50,000 bp in length. Because of the immense size and complexity of the genome, the …
DNA, RNA, and Protein Extraction: The Past and The Present
The extraction of biomolecules, DNA, RNA, and protein, is the most crucial method used in molecular biology [ 1 ]. It is the starting point for downstream processes and product development including diagnostic kits. DNA, RNA, and protein can be isolated from any biological material such as living or conserved tissues, cells, virus particles, or ...
Current Nucleic Acid Extraction Methods and Their …
NAE methods encompass extraction of both DNA and RNA but can be more broadly characterized into chemically driven or solid-phase methods; both contain the four steps mentioned above [1, 4, 5].In the next sections, we will review the working principle of and/or rationale for the main methods used nowadays in the biological and medical …
Evaluation and application of an efficient plant DNA extraction
For DNA extraction from plants, the cetyl trimethylammonium bromide (CTAB) method is one standard way to isolate high ... compared the quality of DNA extracted from tomato leaves by sodium dodecyl sulfate (SDS) extraction and NaOH rapid extraction methods, finding that the ranges of A 260 /A 280 and A 260 /A 230 of isolated …
Sodium Dodecyl Sulfate
Sodium dodecyl sulfate (SDS)-PAGE is a method used for separating and identifying proteins according to their molecular mass. Protein mixtures are boiled in a buffer containing SDS and a reducing agent. ... With respect to STUbL activity, K63-linked chains are known to be involved in DNA damage repair (Galanty et al., 2012; Luo et al., 2012; ...
The Evolution of DNA Extraction Methods
Salting-out method. The salting-out method is a non-toxic DNA extraction method described by Miller, Dykes, and Polesky in 1988. The DNA-containing sample is added to 3mL of lysis buffer (0.4 M NaCl, 10 mM Tris–HCl pH 8.0 and 2 mM EDTA, pH 8.0), SDS and proteinase K. The mixture is incubated at 55–65°С overnight.
Alkaline Lysis Method: How it Works in 5 Simple …
Alkaline lysis was first described by Birnboim and Doly in 1979 and has, with a few modifications, been the preferred method for plasmid DNA extraction from bacteria ever since.[1] The easiest way to …
Nucleic acid protocols: Extraction and optimization
A simplified, semi-unified, protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources. • DNA and RNA are under triple protection …
The Evolution of DNA Extraction Methods
The principal behind DNA extraction consists of the following steps: (1) disruption of cytoplasmic and nuclear membranes; (2) separation and purification of DNA from other components of the cell …
Modified CTAB protocols for high‐molecular‐weight DNA …
Instead of using commercial kits, modified cetyltrimethylammonium bromide (CTAB)‐ or sodium dodecyl sulfate (SDS)‐based methods have been adopted for HMW DNA extraction in various plant groups (Aboul‐Maaty and Oraby, 2019; Li et al., 2020; Jones et al., 2021); however, none of these established protocols have been tested on …
Analytical approaches to differential extraction for sexual assault
Isolation of DNA contributions to link a sample to an alleged offender requires precise chemical treatment of each sample with the goal of separating epithelial cells from non-sperm cells. ... the initial digestion of epithelial cells is performed with proteinase K (Pro-K) and sodium dodecyl sulfate (SDS), leaving sperm cells intact. Adding the ...
Removal of humic substances from soil DNA using aluminium sulfate
Here we report an effective protocol for the removal of humic substance from soil DNA. The protocol involves flocculation of the humic substance by excessive Al (3+), then removal of superfluous Al (3+) via pH adjustment and finally release of soil microbial DNA by SDS lysis. This technique is superior to that employed by the UltraClean Soil ...
A handmade DNA extraction kit using laundry powder; …
Here we describe an in-house kit for high throughput DNA extraction using laundry detergent. A simplified lysis buffer made only from 0.08 M EDTA, 0.1 M Tris, and laundry powder is the core of our protocol. ... Apart from replacing proteinase K and the sodium dodecyl sulfate treatment by the laundry detergent, our protocol instructs a lysis ...
Evaluation of Five Methods for Total DNA Extraction from …
DNA extraction is a routine step in many insect molecular studies. A variety of methods have been used to isolate DNA molecules from insects, and many commercial kits are available. ... The sodium dodecyl sulfate (SDS) and cetyltrimethyl ammonium bromide (CTAB) methods are commonly used for DNA extraction from diverse …
Rapid removal of dextran sulfate sodium from tissue RNA …
1. Introduction. Dextran sulfate sodium (DSS) administration to animals to induce localized, colonic inflammation in vivo is the most widely used experimental model for studying the pathogenesis of inflammatory bowel diseases (IBD) and evaluating novel anti-inflammatory therapeutics. DSS, a negatively charged, sulfated polysaccharide, first …
Precipitation Procedures
Slowly add ammonium sulfate to the desired percent saturation (44) and stir for 10–30 min. Pellet proteins by centrifugation. ... TCA is added to the extract to a final concentration of 10–20% and the proteins are allowed to precipitate on ice for 30 min (46). Alternatively, tissue may be homogenized directly into 10–20% TCA (35, 47).
Preparation of Plasmid DNA by Alkaline Lysis with Sodium …
Abstract. In this protocol, plasmid DNA is isolated from small-scale (1–2 mL) bacterial cultures. Yields vary between 100 and 5 µg of DNA, depending on the copy number of the plasmid. Miniprep DNA is sufficiently pure for use as a substrate or template in many in vitro enzymatic reactions. However, further purification is required if the ...
10% Sodium Dodecyl Sulfate (SDS) (-015) | Rockland
SDS, sodium dodecyl sulfate is an anionic surfactant. SDS is commonly used in laboratory as component of buffer for cell lysis, cell lysis during DNA extraction and mostly in SDS-PAGE running buffer. Indeed, SDS is an anionic detergent applied to protein sample to linearize proteins and to impart a negative charge to linearized proteins.
DNA extraction- Introduction, Methods, History, Types, …
DNA extraction is a fundamental laboratory procedure that involves isolating DNA from cells or tissues, enabling researchers to study and analyze genetic information for a wide range of applications, from medical diagnostics to forensic investigations and genetic research. ... NaCl, sodium lauryl sulfate, and SDS. No …
A Method of DNA Extraction from a Wide Range of …
A Method of DNA Extraction from a Wide Range of Objects via Treatment with Ammonium Salts. BIOLOGICALS TECHNOLOGY. Published: 14 December 2021. …
CTAB or SDS-Based Isolation of Plant's DNA | SpringerLink
Hence, it is a better-preferred detergent during plant DNA extraction. SDS (sodium dodecyl sulfate) is an anionic detergent that denatures the proteins in the biological membranes resulting in their disruption and release of cellular contents. As SDS linearizes the proteins, it also leads to the dissociation of nucleic acid-protein complexes. ...
A modified SDS-based DNA extraction method from raw …
The optimized method was verified by extracting approximately 2020–2444 ng DNA/mg soybean with A 260/280 ratios of 1.862–1.954 from five biotech and non-biotech soybean varieties. Only 0.5 mg of soya was required to obtain enough DNA for PCR amplification using the optimized SDS-based method.
What Is DNA Extraction? Separating Proteins, Membranes
DNA extraction involves isolating deoxyribonucleic acid (DNA) from cells or tissues, a foundational step in molecular biology research. At its core, this process consists of breaking open the cell (lysis), removing proteins and other cellular components, and separating the DNA. This isolated DNA can then be studied, manipulated, or used in ...
Development of an ammonium sulfate DNA extraction method …
We developed an ammonium sulfate DNA extraction method (A) and compared it with two other methods (B and C). DNA quality and quantity, β-globin amplification, and detectability of two cancer associated gene mutations were evaluated. Method A showed the best DNA yield, particularly when the cell number was very low.