Rubicon Genomics – Single Tube Library Preparation Kit for Illumina® Platforms
The first step in any successful sequencing experiment is the preparation of the DNA to be sequenced. In a typical workflow, the DNA is first purified using any of a number of column clean-up methods and then mechanically sheared to yield thousands of short double stranded molecules. At this point, if the DNA is available in less than microgram amounts, a library must be produced. To ensure the highest quality libraries for sequencing, especially when working with limiting (nanogram) quantities of DNA, the library preparation method must be highly efficient, accurate, and not require purification steps where DNA can be lost.
ThruPLEX is a three step process that is performed in a single tube
Step 1. ThruPLEX technology features improved repair chemistry.
Step 2. The background is reduced using double-stranded adaptors with no single-stranded tails. Blunt end ligation occurs with high-efficiency. Blocked 5’ends reduce adaptor-adaptor ligation.
Step 3. Background is reduced by destroying unused adaptors after ligation.
Overall, the time to complete the process is reduced by using compatible buffers and multiplexed enzymatic reactions that do not require intermediate purifications. Continue reading »