The RT primer was designed according to the Yanai protocol23 (link) with an anchored polyT, an 8 nt unique barcode, a 6 nt UMI (unique molecular identifier), the 5’ Illumina adapter (as used in the Illumina small RNA kit) and a T7 promoter. The barcodes were designed with a hamming distance of at least two between them. Primers are desalted at the lowest possible scale, stock solution 1 μg/μL. The oligo sequences can be found in Supplementary Table 3 .
Protocol detail
RT Primer Design for Illumina Sequencing
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Oligo
Primers
Corresponding Organization :
Other organizations : University Medical Center Utrecht, University of California, Santa Barbara
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Variable analysis
independent variables
- RT primer design according to the Yanai protocol
- Anchored polyT
- 8 nt unique barcode
- 6 nt UMI (unique molecular identifier)
- 5' Illumina adapter
- T7 promoter
- Not explicitly mentioned
- Barcodes with a hamming distance of at least two between them
- Primers desalted at the lowest possible scale, stock solution 1 μg/μL
- No positive or negative controls explicitly mentioned
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