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An Introduction to RNA-seq

library prep which includes end-repair, adapter ligation and PCR amplification to give you a library ready for sequencing. Why Bother With Strand Information? There has been a lot of discussion…

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AAV Production Part II: Virus Purification

…Sigma-Aldrich sells iodixanol as ‘Optiprep’. See the paper ‘AAV-mediated gene targeting methods for human cells’ for the different iodixanol solution recipes. First, prepare PBS-MK and NaCl/PBS-MK buffers. PBS-MK buffer: Dissolve…

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Useful Tips to Keep on Top of New Literature

…on what is already in your library and what you’ve recently read. These recommendations are available directly through ReadCube making it simple to add them straight to your library. CiteULike…

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RNAseq Library Preparation: From Cells to cDNA

…(next generation) sequencing of cDNA 5) in silico sequence alignment1, 2. This article is about the two first stages of making an RNAseq library (steps 1- 3). Starting Material Preparation…

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Proteomics and Good Mass Spectrometry Data

…if not removed prior to sample prep. Whew! That was a lot. Maybe most importantly, you should have everything set up for sample prep day. And plan your experiments carefully…

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Nanopore Sequencing: An Update

…and I got an update from multiple users at the UK Genome Science conference. Library prep methods are pretty standard: DNA, frag, end repair, A-tail, adapter ligation. Sequencing accuracy is…

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Next Generation Sequencing: The Three Main Technologies

…(but without the cloning part!). The different NGS platforms have slightly different library preparation methods, the major difference being ‘cluster generation’ for Illumina, and ‘emulsion PCR’ for Roche and Life…

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DNA Extraction from FFPE Tissues for NextGen Sequencing

…nucleic acid crosslinking caused by formalin fixation without causing further DNA damage. Beyond that, extracted DNA requires controlled shearing into manageable fragments for NGS library preparation. DNA Extraction and Shearing…

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Probability Theory and Molecular Barcodes

…during the library preparation, ligate the fragments from each sample to a short, characteristic barcode so that they all share a common sequence. Then sort the reads according to their…

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