Avoid detours! A one-stop solution for key raw materials in NGS library construction is coming!

Figure 1. Conventional DNA library construction process (left) and RNA library construction process (right), taking the Illumina platform as an example

Reverse transcription synthesis is the core part of RNA-Seq, New type of high efficiency reverse transcriptase, which integrates multiple functions such as high sensitivity, high yield of cDNA, and compatibility with templates of complex structures, is a research hotspot. The ZymeEditor™ enzyme modification platform of Yeasen has obtained an all-round reverse transcription product with comprehensively improved performance through directional modification of M-MLV. It can be used in multiple application scenarios such as RNA-seq, single-cell sequencing, cDNA cloning and library construction.

Figure 2. Performance of 13488ES applied in RNA-seq

Limited by the short read length of the sequencing platform, DNA needs to be randomly fragmented before library construction. In the early stage, enzymatic fragmentation was daunting due to problems of uniformity and bias. Yeasen Biotechnology has developed two unique fragmentation enzymes, A vigorously-acting fragmentation enzyme that functions at 37℃ for 3-30 minutes and a mildly - acting fragmentation enzyme that functions at 30℃ for 10-40 minutes. Enzymatic fragmentation products based on random fragmentation are gradually improving their shortcomings, They are combined into modular products for DNA fragmentation, end repair and A-tailing that meet the numerous demands of different samples and different sequencing types.

Figure 3. Fragmentation effects of different DNA fragmentation enzymes: vigorously acting fragmentation enzyme (left) and mildly acting fragmentation enzyme (right)

Library conversion rate is an important indicator for evaluating library quality,A high library conversion rate to some extent means better library richness and better uniformity of sequencing data. Conventional T4 DNA ligase focuses on optimizing high-efficiency ligation, but fragments are prone to self-ligation, which reduces the library conversion rate and affects library richness and sequencing quality. The Yeasen ZymeEditor™ enzyme engineering platform conducts directional modification on T4 DNA Ligase to obtain a new mutant, combined with a carefully optimized ligation buffer,the ligation module product with high thermal stability and a low fragment self-ligation rate has been developed, which significantly improves the library conversion rate.

Figure 4. Thermal stability and linker residue analysis of 12996ES

PCR amplification enzymes are all required in NGS library preparation methods. Most high - fidelity DNA polymerases have 5'→3' polymerase activity and 3'→5' exonuclease activit, It can synthesize DNA in the 5'→3' direction while correcting the erroneously incorporated bases, Thus, it can rapidly and with high fidelity amplify DNA fragments. But few enzymes are specifically designed for NGS, There are not many library amplification products that are efficient, accurate, specific, and able to amplify targets of different sizes and GC contents without bias, and at the same time have the ability to premix primers in advance. Yeasen Biotechnology has developed a highly accurate DNA polymerase with a unique design and an optimized hot start PCR premix,It is specifically designed for efficient, high-fidelity, and low-bias NGS library amplification, addressing the challenges of complex NGS library amplification.

Figure 5. Stability and amplification error rate analysis of 12980ES

In addition to the above series of NGS library construction module product, We also provide one-stop raw enzyme products, to meet the combined needs of different customers. Yeasen's NGS library construction raw material products undergo strict factory quality control standards, strict batch performance and stability quality control, so that you can build libraries without worry.