Molecular Biology Services_Custom Services

Location: Home > Custom Services > Molecular Biology Services

DNA/Gene Synthesis and Purification

Introduction Deoxyribonucleic acid (DNA) synthesis is a process by which copies of nucleic acid strands are made, and till now, Oligonucleotide synthesis is one of the most widely used technology to synthesis relatively short fragments of DNA. Opposite of biosynthesis, the Oligonucleotide synthesis proceeds in the 3’→5’ direction. The method includes four different steps: 1. Detritylation (de-blocking): The synthesis cycle initiated by removing the 5’-DMT protecting group of the solid-support-linked nucleoside, the step results in the solid support-bound oligonucleotide precursor bearing a free 5’-terminal hydroxyl group. 2. Coupling: Once the DMT has been removed, the free 5’-OH of the solid-support-linked nucleoside is able to react with the next nucleotide, which is a phosphoramidite monomer. 3. Capping: The capping step is performed by treating the solid support-bound material with a mixture of acetic anhydride and 1-methylimidazole. This step could eliminate the un-reacted 5’-OH react during the next cycle. 4. Oxidation: The phosphite triester formed in this cycle is unnatural and unstable. Therefore, we treat the support-bound material with iodine...

Details >>
Polypeptide Synthesis, modification and coupling

Polypeptide Synthesis In organic chemistry, peptides can be chemically synthesized by coupling the carboxyl group of one amino acid to the amino group of another amino acid molecule. Solid-phase synthesis and solution-phase synthesis are two major ways of peptide synthesis, and Fmoc solid-phase synthesis is one of the most popular ways of peptide synthesis. Fig. 1 solid phase peptide synthesis. In Fmoc synthesis, the amino acid was attached to an insoluble resin support, this allows the reaction by-products at each step could be removed by simple filtration and washing. The N-terminal of the amino acids were protected by the Fmoc group and coupled to the C-terminal of the growing chain after activation. The Fmoc group then removed by piperidine treatment and the process was repeated till the wanted peptide synthesis finished. After the peptide synthesis process is finished, the peptide is removed from the resin by TFA (trifluoroacetic) treatment. The protecting groups on side chains are removed by the crude linear peptide. After peptide collection, the reverse-phase HPLC is used for the purification of t...

Details >>
RNA/LncRNA/siRNA Synthesis and Purification

Introduction In vitro RNA synthesis requires many factors such as DNA template, RNA polymerase, NTPs and other components. Usually, we use RNA synthesis kits to synthesis RNA in vitro. Similar to DNA, the RNA molecules collect from synthesis could be purified by RNase free-PAGE or RP-HPLC. Why Leading Biology? At Leading Biology, we custom protein purification design for every single protein to ensure the production and recovery rate as high as possible. Working with us, you will get stability, and it means a reliable partner to help streamline your R&D process. Working with us, you will get the guaranteed service to accommodate your requirements. · Vigorous quality control system to ensure the required quality and reproducibility · Competitive price with fast turnaround time Contact Information Please obtain a quote before ordering, and refer to the quote number when you place an order. Orders are typically confirmed within 12 hours. Have a Question? Email us info@leadingbiology.com Order Products: Order Related Products...

Details >>
Nucleic acid extraction

Introduction Nucleic acid extraction plays a vital role in molecular biology since the quality of nucleic acids can affect the performance of downstream reactions, high quality and high yield of nucleic acids are necessary. Nucleic acid extraction including DNA isolation and RNA isolation. DNA Isolation There are four steps to extract DNA from cells or tissues. 1. Break the cell membranes open to expose the DNA (cell lysis) 2. The cell lysates are treated with a concentrated salt solution to make debris clump together. 3. Centrifuge the solution to separate the clumped cellular debris from the nucleic acid. 4. DNA could be isolated by adding ethanol, isopropanol, RNase or even use the minicolumn. RNA Isolation RNA isolation is similar to DNA isolation, only the last step is different. Instead of adding RNase into the precipitate, DNase was added, after purification, isolated RNA molecules were collected. Why Leading Biology? At Leading Biology, we custom protein purification design for every single protein to ensure the production and recovery rate as high as possible. Wo...

Details >>
Molecular Markers

Introduction Molecular Marker is a molecule contained in an organism or non-organism, it could be used to reveal certain characteristics about the source. There are three different molecular markers: genetic markers, biochemical markers and non-biological markers, while the genetic marker is the marker which we gonna discussing below. A genetic marker is a fragment of DNA that is associated with a certain location within the genome, it could correspond to a gene or non-coding regions of the genome, differences at the DNA level could be detected. There are two types of molecular markers: 1. Based on nucleic acid hybridization (non-PCR based approaches): RFLP (Restriction Fragment Length Polymorphism) was the first technology employed for the detection of polymorphism, based on the DNA sequence differences. RFLP is mainly based on the altered restriction enzyme sites, as a result of mutations and recombinations of DNA. Fig.1 RFLP analysis procedure 2. Based on PCR amplification (PCR-based approaches): Including locus non-specific markers like random amplified polymorphic DNA (RAPD) and amplifi...

Details >>

Contact Us

Inquired items

Inquired items

All Departments