Technical Note: The use of molecular barcodes in Anchored Multiplex PCR

MBC Adapterproduct details

Key Points

  1. Sample indexes differ from molecular barcodes (MBCs)
  2. Archer MBCs and downstream error correction
  3. The number of unique MBCs associated with each target region is used to deduce copy number

Sample barcoding, also called sample indexing, is a common approach to labeling samples for multiplex sequencing and analysis. All nucleic acids in a sample are labeled with the same sequence tag, and the resulting library is pooled with other libraries and sequenced in parallel in a single run. Then, during analysis, the sample-specific indexes enable the software to separate the multiplexed sequence data in sample-specific data sets.

Molecular barcoding differs from sample indexing, in that each molecule in a sample is labeled with a unique sequence prior to PCR amplification. With each nucleic acid in the starting material tagged with a unique molecular barcode (MBC), sequence analysis software can filter out duplicate reads and PCR errors to report more accurate unique reads.

There are two major advantages to using molecular barcodes for targeted sequencing:

  • Molecular barcoding exceeds the limited number of unique reads that are identified by alignment-based de-duplication
  • Anchored Multiplex PCR reduces the time spent de-duplicating while providing high quality reads for downstream analysis

Key points from the Molecular Barcodes technical note

Sample indexes differ from molecular barcodes (MBCs)

Sample indexes differ from molecular barcodes (MBCs)

Archer MBCs and downstream error correction

Archer MBCs and downstream error correction

The number of unique MBCs associated with each target region is used to deduce copy number

The number of unique MBCs associated with each target region is used to deduce copy number

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For Research Use Only. Not for use in diagnostic procedures. For Research Use Only. Not for use in diagnostic procedures.