Genomic DNA (gDNA)

We offer genomic DNA (gDNA) from all of our cell lines. Cells are broken down by cell lysis, Proteinase K and RNase A are added, and the gDNA is cleaned over columns. The quantity is determined by absorbance measurement in the NanoDrop. The DNA is stored in TE buffer (10 mM Tris-CL, 0.5mM EDTA pH 9.0). The conditions exclude interference with PCR or other enzymatic reactions in the downstream process.

Fragment length & concentration

The purified DNA has a fragment length of up to 50 kb. The DNA is sold at 5 µg each and the concentration of the solution is indicated on the respective label. We always try to set the concentration to 100 ng/µl. To find out the exact concentration, to order larger quantities of DNA, or if you require a DNA panel for controls, please submit an inquiry.

What is genomic DNA?

Deoxyribonucleic acid (DNA) is a type of organic substance with a very complicated molecular structure that is present in every type of living cell, from bacteria to humans and plants. The genetic information in a person's DNA makes it possible for traits to be passed down through a family.

Genomic DNA was found to be a molecule in 1869, but it wasn't clear what it meant for inheritance until 1943. With the help of biophysicists Rosalind Franklin and Maurice Wilkins, James Watson and Francis Crick in 1953 discovered that DNA had the structure of a double-helix polymer, a spiral made up of two DNA strands twisted around each other. As a result of this discovery, scientists made huge strides in their understanding of DNA replication and the genetic regulation of cellular processes as a result of this discovery.

There is a broad range of gene counts across species. The amount of coding DNA in eukaryotic genomes is very small. The vast majority of eukaryotic genomes are made up of noncoding DNA, which is mostly made up of repetitive sequences. The majority of the function of noncoding DNA is unclear, while some of it is known to have structural and regulatory roles. Ploidy, the number of copies of a given gene in a cell, varies from creature to organism.

What is DNA made of?

DNA is a molecule whose two strands are each made up of a long chain of monomer nucleotides. DNA's nucleotides are made up of the sugar molecule deoxyribose, the phosphate group, and one of four nitrogenous bases: adenine, guanine, cytosine, or thymine.

Covalent connections between the phosphates of two successive nucleotides produce a phosphate-sugar backbone from which the nitrogenous bases extend. Bases form hydrogen bonds with one another to keep one strand attached to the other; this bonding is sequence-dependent, with adenine only connecting to thymine and cytosine only bonding to guanine.

DNA has a very stable structure that allows it to function as a blueprint for making copies of itself and the associated RNA (ribonucleic acid) molecule through a process called transcription. The term "gene" is used to refer to a specific region of DNA that contains instructions for making a certain protein in a cell.

The organization of a DNA strand

DNA may exist in both single- and double-stranded forms with the latter being much more stable. In double-stranded DNA, the nitrogenous bases of one polynucleotide are linked to the nitrogenous bases of another polynucleotide through chemical bonds called hydrogen bonds.

Each A in one strand always couples with a T in the other strand, and each C always pairs with a G; Double-stranded DNA formed in this way resembles a ladder, with sugar phosphates serving as the side supports and base pairs functioning as the rungs.

Please take note that the sugar-phosphate termini of the two polynucleotides that make up double-stranded DNA are anti-parallel, or oriented in opposing directions, since the DNA strands are "upside down" relative to one another. This indicates that the sugar-phosphate chain in one strand is oriented 5' to 3', whereas the other is oriented 3' to 5'. A cell's and an organism's overall functions are regulated by the exact sequence of A, T, C, and G nucleotides in its DNA, which is why it's so important to realize that each creature has its own DNA.

Working with nucleic acid

The DNA molecule is quite stable. However, nucleases should not be added to DNA solutions since they degrade DNA. DNA is made up of extremely big molecules that are extremely brittle. Excessive and harsh pipetting and vortexing should be avoided to protect DNA. Since DNA can be degraded by acid hydrolysis if kept in water, it is best to keep it in TE buffer, pH 7.4.

Genomic DNA applications

CLONING

By digesting genomic DNA with one or more restriction enzymes, and then enzymatically ligating the digested fragments to the linearized vector, the traditional method for cloning DNA fragments into a plasmid or other vector has been established. By using unusual cutters like NotI, large fragments that can be used for YAC and BAC cloning can be made.

Techniques based on polymerase chain reaction (PCR) can also be used to carry out cloning. Typically, these typically involve amplifying DNA fragments with polymerase chain reaction (PCR) before ligating them into recipient vectors, where targeted point mutations can be introduced to modify function.

Sequencing, hybridization mapping, site-directed mutagenesis, and regulation of transcription and translation all rely on cloned products. Other applications include transfection studies to investigate the structure and function of known proteins, as well as the overexpression of particular proteins (with or without tags).

SEQUENCING

DNA may be sequenced once the process of cloning and propagating clones in an appropriate host has been completed. In general, sequencing reactions are exceedingly reliable and almost never fail to yield at least some data. Even though there are a number of factors connected to the structure of DNA sequences that have the potential to influence base calling, total failure of reactions is extremely uncommon.

PCR AMPLIFICATION

PCR is a highly effective method for preparing DNA fragments for other uses including sequencing, cloning, and genotyping. Fluorescent dyes are often added during PCR if automated analysis will be performed afterwards. Five to fifty nanograms (ng) of human genomic DNA is often used in PCR procedures.

GENOTYPING

Genotyping refers to a group of tests that may be used to determine a person's genetic identification and the inheritance patterns of certain characteristics. DNA mutations are associated with diminished protein function, which in turn affects a cell's observable biomarker or phenotype. Strong genotyping tests have applications in forensics, disease gene mapping, pharmacogenomics, evolutionary biology, and the study of population dynamics. Single-base alterations, deletions, insertions, repetitive elements, rearrangements, and methylation are all examples of the types of genetic variation that may occur.