How does the Illumina/Nextera tagmentation enzyme make sure that both p5 and p7 adapters are added?
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The tagmentation enzyme used by Illumina is pre-loaded with both read 1 and read 2 DNA oligo tags in a complex DNA-enzyme structure which makes sure that sequences are added to both ends of the DNA fragmentation site. This is the tagmentation transposome which is used in both bead-linked and free-solution tagmentation enzymes. Subsequent protocol steps are optimized to produce library fragments with one p5 side adapter and one p7 side adapter to allow for sequencing compatibility.
The following kits use a free-solution transposome:
Nextera XT
Illumina Tagment DNA Enzyme and Buffer Kits
The following kits use a bead-linked transposome (the following use different bead-linked versions):
Illumina DNA Prep
Illumina DNA Prep with Enrichment
Illumina DNA PCR-Free
Illumina RNA Prep with Enrichment (tagments double stranded cDNA)
The bead-linked transposome allows for greater flexibly of input DNA amounts so that a wide range of DNA inputs can be used to achieve a consistent library size.
For any feedback or questions regarding this article (Illumina Knowledge Article #5831), contact Illumina Technical Support .
