Protocol detail

Illumina-based MPRA Sequencing Protocol

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Sequencing of 151 nt-long single-end reads was performed on an Illumina MiSeq platform using the MiSeq Reagent Kit v3 150 cycles (Illumina). Fastq files were processed, and the data were analyzed using previously described MPRAdecoder script [93 (link)]. Position weight matrix logos for sequences that dramatically increase (≥10-fold) eGFP mRNA level according to the MPRA screening were defined using the pLogo software [94 (link)].

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Omelina E.S., Letiagina A.E., Boldyreva L.V., Ogienko A.A., Galimova Y.A., Yarinich L.A., Pindyurin A.V, & Andreyeva E.N. (2022). Slight Variations in the Sequence Downstream of the Polyadenylation Signal Significantly Increase Transgene Expression in HEK293T and CHO Cells. International Journal of Molecular Sciences, 23(24), 15485.

Publication 2022

MiSeq platform MiSeq Reagent Kit v3 150 cycles

Mrna

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Variable analysis

independent variables

Sequencing of 151 nt-long single-end reads

dependent variables

EGFP mRNA level

control variables

Illumina MiSeq platform
MiSeq Reagent Kit v3 150 cycles (Illumina)
MPRAdecoder script [93]
PLogo software [94]

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