Why is DNA relatively pure after tac cloning
For multiple molecular biology and genomics applications, from cloning and PCR to genome sequencing and chip analysis, DNA purification is necessary road. However, choosing the best purification kit is not easy. For example, when you are purifying genomic DNA, you need a kit that handles nucleic acids gently without isolating plasmid DNA. Moreover, next-generation sequencing technology has very high requirements on samples, and we also need some specialized products to purify DNA. Here we introduce some new options.
Salt precipitation method
Epicentre's (an Illumina company) MasterPure? Complete DNA Purification Kit uses a patented salt precipitation method. This series of kits covers a variety of sample types, including blood, fixed tissue, plants, yeast, insects, etc., and can bring high yields of nucleic acids in just 30 minutes with minimal loss.
According to Cristine Kinross, senior product manager at Epicentre, with these kits, users do not need to perform additional purification before sample preparation for sequencing or other molecular biology applications. They remove proteins and other cellular debris without introducing toxic compounds that must be removed, thereby eliminating multiple washes and improving nucleic acid yields.
Kinross said: “The excellent purity of recovered nucleic acids using MasterPure allows samples to be directly entered into sequencing library preparation.” Of course, it is also suitable for other applications, including microarrays, qPCR and cloning. In addition, this kit can also be used for RNA purification. After obtaining the total nucleic acid, total RNA can be obtained after DNase treatment.
Variety of kits
Promega offers a wide range of nucleic acid purification kits employing several different strategies. For example, the Wizard? Genomic DNA Purification Kit is based on the classic precipitation method and can extract genomic DNA from whole blood, tissue culture cells, animal and plant tissues, yeast and bacteria, which is flexible and efficient; while the ReliaPrep? series kits use column extraction methods , simple and fast. In addition, Promega also offers magnetic bead-based purification kits.
Eric Vincent, global product manager for genomics at Promega, said: "Although the exact composition of the solution will vary, the basic process of lysis, binding, washing and elution is applicable to different binding methods (such as silica gel). or cellulose), different purification formats (purification columns or magnetic beads), and manual and automated purification operations. "For some very important sample types (such as FFPE samples), some specialized operations or processing are also required.
“We developed unique binding/wash buffers that effectively remove substances that inhibit downstream reactions, and optimized systems allow researchers to capture small amounts of nucleic acids and deliver them in small volumes (less than 30 μl ) elution, we have also developed automated solutions suitable for a variety of reagents and throughputs," said Vincent.
These kits provide a complete solution for elution of nucleic acids in extremely small volumes while avoiding inhibitor residue in the eluate. After purification by these kits, nucleic acids can be used for downstream analysis such as PCR, Sanger sequencing, next-generation sequencing, and chips.