May 16, 2024
Version 3
Nextera XT at 0.2X On the Mantis V.3
- Allen Institute for Brain Science1
- 1Allen Institute
Protocol Citation: Allen Institute for Brain Science 2024. Nextera XT at 0.2X On the Mantis. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk57y1l5r/v3Version created by Allen Institute
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 12, 2021
Last Modified: May 16, 2024
Protocol Integer ID: 99989
Keywords: Nextera XT, Nextera, Formulatrix Mantis, Mantis
Abstract
Protocol to generate Nextera libraries using 0.2x reagents in a 96-well PCR plate using the Formulatrix Mantis instrument.
Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Materials
1.1. Associated Library paper work
1.2. 96-well plate with normalized cDNA at 50 pg/µl (Stored at 4ºC)
1.3. Library plate labels
1.4. 5x Eppendorf twin.tec® 96-well PCR Plates (VWR 47744-106)
1.5. 4x Formulatrix molecular grade HV chips for Mantis instrument (MCHSVM96)
- Filtered pipette tips (GP-L200F, GP-L1000F)
1.7. Nextera XT DNA Sample Preparation Kit for 96 Samples (Illumina FC-131-1096)
1.7.1. Tagment DNA Buffer (TD) (15027866)
1.7.2. Amplicon Tagmentation Mix (ATM) (15031561)
1.7.3. Nextera PCR Master Mix (NPM) (15027920)
1.7.4. Neutralize Tagment Buffer (NT) (15031559)
1.8. Microseal ‘B’ Adhesive seal (BioRad MSB1001)
1.9. VWR Aluminum Foil seals for PCR, Sterile (VWR 60941-076)
1.10. Index primers combined at 2.5 µM each in 96 well plate format.
1.11. Small Kimwipes (VWR 21905-026)