Viral transport media (VTM) is a solution used to preserve and transport viruses, chlamydia, and mycoplasma for research, diagnostic testing, and cell culture. VTM helps to maintain the integrity of RNA viruses, which can be damaged by ultra-low temperature freezers or liquid nitrogen.
VTM are prepared with buffered proteins (serum, albumin, or gelatin), salt solutions and antibiotics. Buffered proteins are used to promote viral growth, provide protection against force during shaking and stirring the medium. A balanced salt solution is added to maintain viable state of the cells, pH and osmotic pressure. Antibiotics are often added to viral transport media to inhibit the development of contaminating bacteria and fungus.
RNA (ribonucleic acid) is a molecule similar to DNA present in the cells for coding, decoding, regulation and expression of genes as protein synthesis and in some viruses act as their genetic material. RNAs are single stranded. RNA covert the message/code provided by the DNA into proteins for cellular activities (development, cellular differentiation, and changing environments) by translation.
RNA purification can be used for molecular analyses such as RT-PCR, cloning, in-vitro translation and transcription, transcriptome analysis, RNA structural and functional studies, depletion, and RNA sequencing sampling preparation. It is also essential for the development of diagnostics and drugs.
RNA extraction is the process of separating RNA from cells present in any biological sample. Mission is to separate pure RNA. The extracted RNA should be of good quality and in efficient amount. Quality should be determined by purity of RNA without contamination such as DNA, lipids and proteins. Avoiding things like detergents, temperature, bad handling, ribonuclease enzyme etc. which can denature RNA so should go through a certain protocol. RNA can be extracted by manual method as well as commercially available kits.
In RNA extraction and purification, Firstly disrupt the cells by adding guanidium thiocyanate and a reducing agent to the sample and then subject it to vigorous shaking or vortexing. This step will also break the disulphide bonds and inactivate the contaminant proteins present in the sample. After cell lysis, add phenol and chloroform-isoamyl alcohol to separate RNA sample from the solution. The aqueous phase contains RNA which is transferred to a separate tube, add isopropanol and centrifuge the solution to precipitate the RNA. Washing the precipitate with 75% ethanol will then remove any impurities from the sample.
DNA (deoxyribonucleic acid) in every living cells are responsible for overall regulation of that cell (growth, survive and reproduce). On certain signals or stage of its life cycle the DNA gives messages to cell by transcription and translation. Each individual of certain species differs from each other on genetic basis and DNA is that genetic material in almost every living organism except of few viruses having RNA as genetic material.
DNA purification can be used for molecular analyses such as PCR, electrophoresis, sequencing genomes, fingerprinting, genetic engineering and cloning. It plays an important role in understanding the genetic cause of disease, detection of virus and bacteria. It is also essential for the development of diagnostics and drugs.
DNA extraction is the process of separating DNA from cells present in any biological sample. Mission is to separate DNA in the nucleus from other cellular components found in the cells. The extracted DNA should be of good quality and in efficient amount. Quality should be determined by purity of DNA without contamination such as RNA and proteins. Avoiding things like detergents, temperature, bad handling etc. which can denature DNA so should go through a certain protocol. DNA can be extracted by manual method as well as commercially available kits.
Step 1. Breaking cells open to release the DNA
The cells in a sample are separated from each other, such as by physical methods like vertexing, and put into a solution containing salt. The positively charged sodium ions in the salt protect the negatively charged phosphate groups that run along the backbone of the DNA.
A detergent is then added. The detergent breaks down the lipids in the cell membrane and nuclei. Hence DNA is released as when these membranes are disrupted.
Step 2. Separating DNA from proteins and other cellular debris
Often a protease is added to degrade DNA-associated proteins.
Step 3. Precipitating the DNA with an alcohol
Ice-cold ethanol or isopropanol added to the DNA sample. DNA is soluble in water but insoluble in salt and alcohol. So by stirring the alcohol layer with a sterile pipette, the precipitate is visible and can be taken out. If there is lots of DNA, you may see a white precipitate.
Step 4. Cleaning the DNA
The DNA sample can now be further purified. It is then resuspended in a slightly alkaline buffer.
Step 5. Confirming the presence and quality of the DNA Optical density readings taken by a spectrophotometer can be used to find out the concentration and purity of DNA in a sample. Also, gel electrophoresis can be used to show the presence of DNA in the sample.
Viral RNA purification kit
GJC Viral RNA purification kits and reagents are available from multiple suppliers. These ready-to-use tools are designed for the streamlined and effective extraction of RNA from viral particles. Some kits are tailored for a particular sample source, such as blood, tissue, or swab.
DNA purification kit
GJC DNA Purification Kit uses a membrane-based system to quickly purify high-quality genomic DNA that is suitable for downstream applications such as agarose gel electrophoresis, restriction enzyme digestion, and PCR analysis. Genomic DNA can be purified from cells in as little as 20 minutes.
VTM (Viral Transport Medium)
The diagnosis of COVID-19 viral infection by culture relies on the collection of proper specimens and proper care to protect the virus in the specimen from environmental damage and the use of an adequate transport system to maintain virus activity. GJC Viral Transport Media carries a specially formulated medium for the transportation of viral specimens. It is designed to maintain the optimum viability and virulence of the viral specimen. GJC VTM allows the safe transfer of viruses for diagnostic purposes. Designed with safety, reliability, and convenience to meet your viral testing needs.
PCR cleanup kit
GJC PCR Clean-Up Kit is designed for rapid purification of single-stranded or double-stranded PCR amplification products (100 bp to 10 kb) from other components in the reaction, such as excess primers, nucleotides, DNA polymerase, oil and salts. This kit combines the advantages of silica binding with a convenient spin-column format, eliminating the need for expensive resins or toxic organic compounds such as phenol and chloroform.