Protocol for TotalScriptâ¢ RNA-Seq Kit

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TotalScript RNA-Seq Kit 7. Appendix Appendix A. Bioanalyzer Profile Of A TotalScript RNA-Seq Library A TotalScript RNA-Seq library was made from 5 ng of random primed Universal Human Reference RNA (UHR) using the TotalScript Optimized Buffer as described in Part 3.A of the procedure. Appendix B. It is recommended to use the Control RNA as a positive control for library prep to determine that all the steps of the protocol are followed as directed. 1. Thaw the Control RNA and keep on ice. Remove 1 ul of the Control RNA and place in a fresh RNase-free tube, then add 99 ul of Nuclease-free water and mix well. Keep diluted Control RNA on ice. 2. Set up the following reaction, as per step 3.A.1. of the TotalScript cDNA Synthesis procedure: 2 μl 1:100 diluted Control RNA 2.5 μl TotalScript First-Strand Buffer 1 μl Random Hexamer Primers 12.5 μl Nuclease-free water 18.0 μl Total 3. Place the samples in a thermocycler with a heated lid. Heat for 2 minutes at 65°C, then hold at 4°C. 4. Proceed to Part 3.B, 1st Strand cDNA synthesis. 5. At Step 4.E. PCR Amplification: a. Amplify the sample for 12 cycles and purify the reaction as directed in Part 4.F. of the protocol. b. Run 1 ul on a bioanalyzer using a high sensitivity chip. 10 www.epicentre.com
TotalScript RNA-Seq Kit Appendix C. TotalScript Index Primer Sequences Index 1 5′-TAAGGCGA-3′ Index 2 CGTACTAG Index 3 AGGCAGAA Index 4 TCCTGAGC Index 5 GGACTCCT Index 6 TAGGCATG Index 7 CTCTCTAC Index 8 CAGAGAGG Index 9 GCTACGCT Index 10 CGAGGCTG Index 11 AAGAGGCA Index 12 GTAGAGGA Visit our technical blog: epicentral.blogspot.com Limited Use Label License: This product and its use are the subject of one or more issued and/or pending U.S. and foreign patent applications owned by Max Planck Gesellschaft, exclusively licensed to New England Biolabs, Inc. and sublicensed to Illumina, Inc. The purchase of this product from Illumina, Inc., its affiliates, or its authorized resellers and distributors conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product by the buyer (whether the buyer is an academic or for profit entity). The purchase of this product does not convey a license under any claims in the foregoing patents or patent applications direct to producing the product. The buyer cannot sell or otherwise transfer this product or its components to a third party or otherwise use the product for the following COMMERICAL PURPOSES: (I) use of the product or its components in manufacturing; or (2) use of the product or its components for therapeutic or prophylactic purposes in human or animals. techhelp@epicentre.com • (800) 284-8474 11
Page 1 and 2: TotalScript RNA-Seq Kit Cat. No. TS
Page 3 and 4: TotalScript RNA-Seq Kit Required Re
Page 5 and 6: TotalScript RNA-Seq Kit Important!
Page 7 and 8: TotalScript RNA-Seq Kit 4.A. Tagmen
Page 9: TotalScript RNA-Seq Kit 4.F. Purify

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Protocol for TotalScriptâ¢ RNA-Seq Kit