This allows for seamless generation of Illumina sequencing libraries using a simple PCR reaction, without the need to purify RNA as in ChRIP-seq, APEX-seq and TRIBE. The lack of purification steps makes RT&Tag adaptable for automation as was done with AutoCUT&Tag32. Together with low cell ...
Reverse transcription polymerase chain reactionWhereas techniques to map chromatin-bound proteins are well developed, mapping chromatin-associated RNAs remains a challenge. Here, we describe Reverse Transcribe and Tagment (RT&Tag), in which RNAs associated with a chromatin epitope are targeted by an ...
We are presenting a new bacterial DNA enrichment method for Oxford Nanopore sequencing called ON-rep-seq. The method exploits an optimized version of Rep-PCR for reproducible amplification followed by a dual-stage Rep-PCR-2 step allowing tagmentation of up to 96 samples in one reaction. Furtherm...
The library amplification reaction was again prepared with the Echo. The Nextera XT Indexes were presented on a 384PP 2.0 plate and transferred to the Library preparation plate so that each well received one of the 384 possible unique index combinations. After that, 1500 nl of Nextera PCR Mas...
In vitro transposition with Nextera Transposomes simultaneously fragments and covalently tags the target DNA, combining these three distinct steps into a single reaction. Platform-specific adapters can be added, and the sample can be enriched and barcoded using limited-cycle polymerase chain reaction ...
Acoustic liquid transfer (Echo 525) was employed to reduce reaction volumes and the number of tips required for a Nextera XT library preparation. Sequencing was performed on an Illumina MiSeq. The final version of Mini-XT has been used to sequence 4384 SARS-CoV-2 samples from N. Ireland ...