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[DESCRIPTION]
This protocol is a group of the following 5 methods from the NEB website:<br />1.&nbsp;<a href="https://www.neb.com/protocols/2012/11/15/nebnext-end-prep-e7370" target="_blank">NEBNext End Prep </a> <br />2. <a href="https://www.neb.com/protocols/2012/11/15/adaptor-ligation-e7370" target="_blank">Adaptor Ligation</a><br />3.&nbsp;<a href="https://www.neb.com/protocols/2012/11/15/size-selection-of-adaptor-ligated-dna-e7370" target="_blank">Size Selection of Adaptor-ligated DNA</a><br />4.&nbsp;<a href="https://www.neb.com/protocols/2012/11/19/pcr-amplification-e7370" target="_blank">PCR Amplification</a><br />5.&nbsp;<a href="https://www.neb.com/protocols/2012/11/19/cleanup-of-pcr-amplification-e7370" target="_blank">Cleanup of PCR Amplification</a>

[GUIDELINES]
The NEBNext Ultra DNA Library Prep Kit for Illumina contains enzymes and buffers that are ideal to convert a small amount of DNA input into indexed libraries for next-generation sequencing on the Illumina platform (Illumina, Inc).&nbsp; <br /> <br /><strong>Adaptor Ligation:</strong><br /><br />If DNA input is &lt; 100 ng, dilute the NEBNext Adaptor for Illumina* 1:10 in sterile water use immediately to a final concentration of 1.5 &mu;M.<br /><br /><strong>Size Selection:<br /></strong><br />This protocol is suitable for input amounts of 50ng or higher. For lower input amounts we recommend that the protocol without size selection be followed.<br /><br />This size selection is for libraries with 200 bp inserts only and&nbsp;is based on a starting volume of 100 &micro;l. For libraries with different size fragment inserts, refer to Table 1 for&nbsp; the appropriate volume of beads to be added.&nbsp;<br /><br />Table 1: Recommended conditions for bead based size selection.<strong>&nbsp;</strong><br /><table><tbody><tr><td rowspan='2' valign='middle'><strong>LIBRARY</strong><br /><strong>PARAMETERS</strong></td><td valign='middle'><strong>APPROXIMATE</strong><br /><strong>INSERT SIZE</strong></td><td><strong>150 bp</strong></td><td><strong>200 bp</strong></td><td><strong>250 bp</strong></td><td><strong>300-400 bp</strong></td><td><strong>400-500 bp</strong></td><td><strong>500-700 bp</strong></td></tr><tr><td valign='middle'>Total Library Size (insert + adaptor)</td><td>270 bp</td><td>320 bp</td><td>400 bp</td><td>400-500 bp</td><td>500-600 bp</td><td>600-800 bp</td></tr><tr><td rowspan='2' valign='middle'><strong>VOLUME TO</strong><br /><strong>BE ADDED (&mu;l)</strong></td><td>1st Bead Selection</td><td>65</td><td>55</td><td>45</td><td>40</td><td>35</td><td>30</td></tr><tr><td>2nd Bead Selection</td><td>25</td><td>25</td><td>25</td><td>20</td><td>15</td><td>15</td></tr></tbody></table><br /><strong>Note: Be sure not to transfer any beads. Trace amounts of bead carry over may affect the optimal performance of the polymerase used in the NEBNext High-Fidelity 2X PCR Master Mix in the subsequent PCR step.<br /><br /></strong>Figure 1: Examples of libraries prepared with human IMR-90 gDNA.<strong><br /><img src='http://i.imgur.com/Jg1AiWp.png' alt='' width='700' height='493' /><br /><img src='http://i.imgur.com/Ytnm7tX.png' alt='' width='700' height='512' /><br /></strong>

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