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Cellecta, Inc.
NGS Prep Kit for sgRNA, shRNA, and DNA Barcode Libraries
Table of Contents
Cellecta, Inc.
  • NGS Prep Kit for sgRNA, shRNA, and DNA Barcode Libraries
GrabGrab
GrabGrab
CELLECTA User Manual
  • 1. NGS Prep Kit for sgRNA, shRNA, and DNA Barcode Libraries
  • 2. NGS Prep Kit Required Materials
    • 2.1. Materials Provided
    • 2.2. Additional Materials Required
    • 2.3. Related Products and Services from Cellecta
  • 3. Procedure Overview
    • 3.1. Genomic DNA Extraction
      • 3.1.1. Large-Scale Genomic DNA Extraction Protocol
    • 3.2. Amplification of sgRNA or Barcode Sequences from Genomic DNA
    • 3.3. NGS Sample Purification
    • 3.4. Sequencing sgRNA Inserts or Barcodes
    • 3.5. Demultiplexing and Aligning Sequencing Reads
  • 4. Troubleshooting
    • 4.1. No PCR Product
    • 4.2. No sgRNA/Barcodes Present in NGS Results
    • 4.3. NGS Data Includes Significant Percentage of Reads from a Single Sequence
    • 4.4. Incorrect Band Pattern on Agilent Bioanalyzer
  • 5. Technical Support
  • 6. Contact Us
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4.3. NGS Data Includes Significant Percentage of Reads from a Single Sequence

Possible reason #1:

Cells used in screen may carry foreign DNA which contain NGS primer binding sites.

Solution:

Do not use cells for your screen which contain foreign DNA with NGS binding sites.

Last modified: 10 May 2018
4.2. No sgRNA/Barcodes Present in NGS Results
4.4. Incorrect Band Pattern on Agilent Bioanalyzer

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