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Today, we are going to show you how to make DNA libraries using the NEBNext Ultra II DNA Library Prep Kit for Illumina. Before you begin, youamp;#39;ll want to make sure that you have available all of the required reagents and equipment that are not included in the kit. Your DNA should be fragmented to the size range suitable for your sample and sequencing requirements. This can be done using either mechanical methods or enzymatic methods. The volume of your sample should be adjusted to 50 ul, and this should contain 500 pg to 1 ug of fragmented DNA. Although the Ultra II kit can be used with very low input amounts, we recommend using higher amounts if available, to minimize the number of PCR cycles required. When working with the reagents included in the kit, itamp;#39;s important to ensure that they are well mixed before use. The first steps in library construction are End Repair and dA-Tailing. In this step, the input DNA is blunted and phosphorylated, and an A is added to the 3
