The SageHLS system is a new platform released by Sage in the United States in 2017. It can rapidly purify high molecular weight DNA directly from cell samples. Chris Boles, Sage's Chief Scientific Officer, has been helping the R&D team develop the SageHLS platform (HLS is an acronym for High Molecular DNA Library Preparation Systems). So, what is the role of high molecular weight DNA in life sciences and biomedical research? How does the SageHLS system work? What new development applications will follow? Let's listen to Chris's introduction to the product.
1. Q: Why are high molecular weight DNA fragments important?
A: The use of extremely long DNA fragments has become a lost art in the life sciences. Back to the early Human Genome Project, because each laboratory uses techniques such as recombinant chromosomes, plasmids, and Southern blotting, it is necessary to consider the use of extremely long DNA fragments. But with the advent of PCR technology in the early 1990s and the advent of short-reading NGS technology, scientists no longer need high-molecular-weight DNA for scientific research purposes. Now, researchers are addressing highly overlapping regions of the genome, large fragment structural variations, and large fragment conformation problems, so high-quality, high-molecular-weight DNA is once again becoming very important.
A: The use of extremely long DNA fragments has become a lost art in the life sciences. Back to the early Human Genome Project, because each laboratory uses techniques such as recombinant chromosomes, plasmids, and Southern blotting, it is necessary to consider the use of extremely long DNA fragments. But with the advent of PCR technology in the early 1990s and the advent of short-reading NGS technology, scientists no longer need high-molecular-weight DNA for scientific research purposes. Now, researchers are addressing highly overlapping regions of the genome, large fragment structural variations, and large fragment conformation problems, so high-quality, high-molecular-weight DNA is once again becoming very important.
2. Q: What long-segment genome technology can the SageHLS platform be used in conjunction with?
A: All systems that require hundreds of Kb to several Mb DNA samples, such as long read length sequencing systems from PacBio or Oxford Nanopore, genomic optical mapping systems from Bionano Genomics or Genomic Vision, and extensive linkage analysis techniques from 10x Genomics or Dovetail Genomics . The DNA quality and fragment size of all of these systems can be improved with the SageHLS platform.
3. Q: What kind of pretreatment does the sample need before SageHLS is loaded?
A: Generally speaking, preprocessing is simple. The sample types we initially focused on had two conditions, 1) important for biomedical research, and 2) working well in the SageHLS system. These samples include whole blood leukocytes, tissue culture cells or bacterial cultures. For these sample types, only a simple centrifugation step is required to wash the cells and resuspend in the loading buffer. The reagents supplied with the instrument will be lysed and purified, and the high molecular weight DNA will not be mixed or broken throughout the process.
A: Generally speaking, preprocessing is simple. The sample types we initially focused on had two conditions, 1) important for biomedical research, and 2) working well in the SageHLS system. These samples include whole blood leukocytes, tissue culture cells or bacterial cultures. For these sample types, only a simple centrifugation step is required to wash the cells and resuspend in the loading buffer. The reagents supplied with the instrument will be lysed and purified, and the high molecular weight DNA will not be mixed or broken throughout the process.
4. Q: How does the new system work?
A: First, the user moves their sample into the gel, and then the platform automatically performs cell lysis and contaminant removal. This process is very fast, and Mb-sized DNA is left in the agarose. Next, the system cleaves the DNA slightly with a non-specific nuclease and recovers the DNA fragment of interest from the gel by an automated elution process.
A: First, the user moves their sample into the gel, and then the platform automatically performs cell lysis and contaminant removal. This process is very fast, and Mb-sized DNA is left in the agarose. Next, the system cleaves the DNA slightly with a non-specific nuclease and recovers the DNA fragment of interest from the gel by an automated elution process.
5. Q: What results did you get from the SageHLS platform?
A: Under normal conditions, we have purified and recovered 200Kb-2Mb DNA fragments in mammalian leukocytes and tissue culture cells. We recovered 1-3 ug of high molecular weight DNA from loading cells containing approximately 10 ug of DNA (approximately 1.5 million human cells), which is sufficient for large sample applications such as genomic optical profiling.
A: Under normal conditions, we have purified and recovered 200Kb-2Mb DNA fragments in mammalian leukocytes and tissue culture cells. We recovered 1-3 ug of high molecular weight DNA from loading cells containing approximately 10 ug of DNA (approximately 1.5 million human cells), which is sufficient for large sample applications such as genomic optical profiling.
6. Q: You are already developing new uses for the SageHLS platform. Can you give me some information?
A: The next step we developed is the technique called HLS-CATCH (HLS-based assisted targeted chromosomal fragment cloning), which uses the CRISPR/Cas9 gene editing technology to target and excise and isolate the genome of interest on HLS. Fragment. We are also investigating several methods for preparing NGS libraries directly in HLS preset cartridges so that we can integrate DNA extraction with NGS library construction. It is very interesting to learn from customers what they want to do with the system.
A: The next step we developed is the technique called HLS-CATCH (HLS-based assisted targeted chromosomal fragment cloning), which uses the CRISPR/Cas9 gene editing technology to target and excise and isolate the genome of interest on HLS. Fragment. We are also investigating several methods for preparing NGS libraries directly in HLS preset cartridges so that we can integrate DNA extraction with NGS library construction. It is very interesting to learn from customers what they want to do with the system.
From Wenjun Jiang et al., 2015, Nature communication
198 kb DNA fragment extracted from E. coli protoplasts based on the HLS platform (treated with CRISPR/cas9 nuclease)
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Huanya Biotech Co., Ltd. is the general agent of Sage Company in China. It has been paying attention to the introduction and development of sequencing related equipment in China. It is committed to assist domestic customers to complete the preparation of sequencing samples better and faster, and achieve higher efficiency. Easier workflow. Company website:
Huanya Biotech Co., Ltd. is the general agent of Sage Company in China. It has been paying attention to the introduction and development of sequencing related equipment in China. It is committed to assist domestic customers to complete the preparation of sequencing samples better and faster, and achieve higher efficiency. Easier workflow. Company website:
Can be used directly to maintain the integrity of cell morphology Guidelines recommend cell preservation fluids for sample preservation and transport. It can be used for sputum, bronchoscopic alveolar lavage fluid, pleural fluid and other liquid samples.
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