UMI

Unique Molecular Identifiers

Unique molecular identifiers (UMI) are molecular tags that are used to detect and quantify unique mRNA transcripts (Kivioja et al., 2012). In this method, mRNA libraries are generated by fragmentation and reverse-transcribed to cDNA. Oligo(dT) primers with specific sequencing linkers are added to the cDNA. Another sequencing linker with a 10 bp random label and an index sequence is added to the 5′ end of the template, which is amplified and sequenced. Sequencing allows for high-resolution reads, enabling accurate detection of true variants.

Advantages:

  • Can sequence unique mRNA transcripts
  • Can detect transcripts occurring at low frequencies
  • Transcripts can be quantified based on sequencing reads specific to each barcode
  • Can be applied to multiple platforms to karyotype chromosomes

Disadvantages:

  • Targets smaller than 500 bp are preferentially amplified by polymerases during PCR


Reagents:

Illumina Library prep and Array Kit Selector



Reviews:

This method has been widely integrated into various sequencing techniques due to its high versatility.

References:

This method has been widely integrated into various sequencing techniques due to its high versatility.