We can provide some common guidelines for primer design and amplification of oligo pools. But for specific experiments, our guidelines can only be used as general guidelines. You need to adjust the specific experimental steps according to the conditions. The guiding principles for the use of oligo pools mainly include the following points:
A set of universal primers will be used before amplification. PCR amplification can achieve the effect of purifying oligo pools because the failed primer sequence (the primer sequence missing at 5 'end) cannot be amplified.
To achieve a higher annealing temperature, we suggest using a high Tm value or longer primers. Higher annealing temperatures can reduce primer mismatch and enhance the specificity of amplification.
In PCR amplification, we recommend not to use conventional Taq polymerase, but to use hot start enzymes, such as Fusion, Q5, Pfu, Vent, Platinum Taq Polymerase, and PrimeTaq™ HotStart Direct PCR DNA Polymerase. The selection of enzymes should be cautious, and some enzymes need to be tested before use.
Due to the different sequence lengths, primer designs, and polymerase systems of different users, the usage of starting template is also different. For PCR templates, it is suggested to use 1/100, 1/400, 1/1600, 1/6400, and 1/25600 of the total reaction volume for gradient test. Set the number of PCR cycles, such as 25 cycles to amplify, and use gel or Bioanalyzer to detect the amount of PCR products. As long as the quantity of PCR products meets the requirements, the smaller amount should be chosen as the final amount of the template.
If multiple sequences need to be amplified from the product, nested PCR is used to amplify all the sequences, and then internal primers are used to amplify each group of sequences. Only one round of PCR may not be enough for getting a good result.
The basic strategy of primer removal depends on the specific needs of downstream application of oligo pools, such as blunt end or sticky end.
For normal cloning, users can insert the restriction sites recognized by type IIS restriction enzymes into the universal primers and then remove the universal primers after PCR amplification. These restriction enzymes can digest near the recognition site to maintain the integrity of variable sequences. After the digestion, the oligo sequences can be inserted into the vector.
Only for research and not intended for treatment of humans or animals
DNA synthesis is a technology that links deoxynucleic acids (adenine, thymine, cytosine, and guanine) together to form DNA. As the cornerstone of modern molecular biology, DNA synthesis plays a pivotal role in the field of synthetic biology. In addition to standard oligos synthesis, we also provide scientific research services such as long oligos synthesis, phosphorothioate oligos (S-Oligo) synthesis, modified oligos synthesis, fluorescent oligos synthesis, and real-time quantitative PCR probes to meet your different needs. Besides, we have been providing high-quality DNA synthesis products (phosphoramidites, controlled pore glass, molecular sieves, etc), and the products have been successfully applied in various fields of molecular biology.
Peptides are synthesized by the condensation reaction of the carboxyl group of one amino acid with the amino group of another amino acid and are widely used in various fields such as antibody preparation, drug development, and polypeptide vaccine development. Each of our polypeptides is accompanied by reliable HPLC and mass spectrometry data, detailed synthesis reports are provided, and the products are sent in a lyophilized state. Experienced staff can assist users in designing peptide chains and make appropriate recommendations for different needs of users, such as antibodies, special markers, large-scale synthesis, etc. In addition, we also offer Custom PNA Synthesis with high quality.
A peptide library is a new technique for studying the structure-function relationship for a protein. A peptide library contains a great number of peptides that have a systematic combination of amino acids. The peptide library provides a powerful tool for drug design, protein-protein interactions, and other biochemical as well as pharmaceutical applications. It also has wide applications in drug screening, target validation, epitope mapping, and vaccine development.
Gene synthesis is a technology that synthesizes genes by artificial methods, which is one of the means of gene acquisition. Compared with the acquisition of genes from existing organisms, gene synthesis does not need templates and is therefore not limited by the source of genes. We use unique gene synthesis design software, which includes a full set of tools to design ideal structural units, thus enabling rapid and efficient gene construction and synthesis in a single reaction. Please do not be limited by restriction sites and polylinkers, we will synthesize the various gene sequences you need.
Polymerase chain reaction (PCR) is a widely used method in molecular biology, which can rapidly replicate millions to billions of specific DNA samples, enabling scientists to extract only a small amount of DNA samples for detailed research. We provide a variety of DNA polymerases (Taq, Bst, Pfu) and corresponding PCR premixes, covering a wide range of scenarios such as high fidelity, high specificity, and rapid amplification. High-quality dNTPs and NTPs (set, mix) are also supplied.
Ribonucleic acid (RNA), as a key material for genetic information transmission and cell regulation, has been extensively studied in molecular biology. Like DNA, RNA is assembled in the form of nucleotide chains; but unlike DNA, RNA exists in nature in the form of single-stranded folds rather than paired double strands. We provide RNasin (RNase inhibitor) and M-MLV reverse transcriptase to provide a complete raw material solution for RNA research. miAnalysis™ series are designed for microRNA quantitative research. And VirusMag™ series is designed for the isolation of viral RNA/DNA or bacterial DNA.
At SBS Genetech, we are at the forefront of offering solutions for isothermal amplification based on our world-class platform. Our Bst DNA/RNA Polymerase is at the core of this platform, which is a mixture of Bst polymerase and extremely thermostable reverse transcriptase. Based on this special enzyme, we have developed PrimeIampTM lyophilized isothermal amplification microbeads series. These series are ready-to-use master mixes, which can perform isothermal amplification directly when the templates and primers are added. With freeze-drying technology, these master mixes are lyophilized into solid microbeads, which can be transported and stored at room temperature with great convenience.
RNA silencing technology has become a powerful tool to study gene function. The success of any RNA experiment depends on high-quality siRNA and effective transfection reagent. With chemical modification, our chemically modified siRNA has much higher stability than the common siRNA. The chemical modification not only enhances the life span of siRNA in serum and cell culture but also enhances its activity in vitro. As for transfection reagent, our ready-to-use siRNA transfection reagent, Sirnafectamine, can be used for a wide range of cell lines, with minimal cytotoxicity and the best cell state after transfection. As Sirnafectamine will protect RNA during the whole process, a very low concentration of siRNA can produce high gene silencing efficiency.
Nucleic acid purification is an important component of molecular biology and has a wide range of applications in medicine and biological sciences. Our nucleic acid purification kit uses first-class silica gel column technology (SiMax™ Spin Column) and magnetic beads technology (VSep™ Magnetic Separators, VirusMag™ One-Step DNA/RNA Isolation Kit, VirusMag™ DNA/RNA Isolation Kit), which can purify DNA/RNA from various sources quickly and reliably. The purity of DNA/RNA purified by our kit is very high and it is suitable for many downstream applications such as sequence determination, cloning, and cleavage.
DNA markers are used to determine the approximate size of molecules on the gel during electrophoresis, based on the principle that the molecular weight is inversely proportional to the mobility through the gel matrix. We provide abundant DNA molecular weight standards that can be used for various fragment lengths. Our fragments of DNA markers have been purified separately by proprietary technology, so their quality is superior to industry standards.
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria and archaea. The gene-editing technology based on this system has a wide variety of applications. Here, we provide various Cas nucleases, synthetic sgRNAs, and T7 Endonuclease I of high quality to improve the accuracy and efficiency of your experiments.
Gene manipulation is a process that uses biotechnology to manipulate genes directly to generate new DNA and has been widely used in research, medicine, industrial biotechnology, and agriculture. Our Premium™ Master Assembly Mix and Topo Cloning kits (pBM23, pBM16A, pBM16K) provide efficient solutions for these types of demands.