Search results for: library%2520prep
Get Prepped: Nanopore Library Preparation Optimization
…and Nanodrop measurements. Nanopore library quality mostly depends on the individual researcher’s technical skills and pipetting technique. In this article, we discuss some pointers to perform a successful NGS library…
Read MoreWGS Workflow: From Sample Collection to Data Interpretation
…quality of the DNA sample. Construct a Library Preparing a DNA library from the genomic sample at hand is a crucial step in the WGS workflow. Essentially, a DNA library…
Read MoreLevel Up Your Drug Screening With CRISPR
…pooled versus arrayed approaches. © 2015 Agrotis and Ketteler [5], licensed under CC BY 4.0. MOI, multiplicity of infection; sgRNA, single guide RNA. Pooled Library Screens A pooled library can…
Read MoreHow To Get Organized With Reference Managers for Science—Mendeley
…to your Mendeley library. This saves time and means that your library is effortlessly maintained and that you won’t lose that one vital paper that may hold the key to…
Read MoreA Primer on Phage Display Libraries
…In a semi-synthetic library, these framework regions are amplified and cloned into the phage display library from B-cells, in differing combinations to achieve different goals. In a synthetic library, these…
Read More8 Approaches to Random Mutagenesis
…then randomly introduce mutations into the gene sequence to create a “library” containing thousands of versions of the gene. Each version (or “variant”) of the gene in the library would…
Read MoreTop 10 Tips for NGS Library Preparation
Making a Next Generation Sequencing (NGS) library can seem a bit daunting to the new user, as failures can be expensive. But don’t be put off, as NGS library preparation…
Read MoreHow to Improve Your WGS DNA Library
…Library Prep Kit for Illumina Systems and Invitrogen™ Collibri™ PS DNA Library Prep Kit for Illumina Systems, it is possible to reduce these biases. References Macintyre G, Yistra B, and…
Read MoreHow To Troubleshoot Your DNA Libraries For 454 Next Generation Sequencing- An Essential Guide.
For those of you who prepare your own DNA libraries, this article will cover the most critical aspects of library preparation to ensure a successful sequencing run. Previous Bite Size…
Read MoreZotero Reference Manager: A Super Simple Guide for Busy Researchers
…library of publications, collaborate easily with colleagues, and seamlessly insert citations in your chosen word processor. Maybe even more importantly, gone are the days of manually writing and formatting your…
Read MoreFrom the Lab to the Library…
…did indeed go back to school to get my Masters in Library Science (MLS).Then I looked at library and university online job sites to look for openings (check out the…
Read MoreHow To Get Organized With Reference Managers for Science—ReadCube Papers
…such as: library generation for your references; cloud syncing of your library; access across devices; and the ability to make notes, highlight text, and annotate PDFs. In addition, the sleek…
Read MoreHow to Access Journal Articles Behind Paywalls
…library has the right issue of the journal. Then, call your institution’s or city’s library and arrange for a loan from that other library. This may cost a nominal fee…
Read MoreThe Recipe for Successful Whole Genome Sequencing
…to obtaining high-quality WGS results and is available exclusively from the Invitrogen™ Collibri™ PS DNA Library Prep Kit and the Invitrogen Collibri PCR-Free PS DNA Library Prep Kit for Illumina…
Read MoreQuantifying Your NGS Libraries
…very high-throughput. Illumina recommends the very capable KAPA library quantification kit, but there are alternatives including Agilent, Qiagen and QuantBio, among others. The KAPA kit is a SYBR-green assay that…
Read MoreThe Cell: An Image Library – An Overview of an Award-winning Multimedia Site
Much more than just an archive, The Cell: An Image Library-CCDB (Cell Centered Database; ‘The Cell’) serves many additional purposes. Whilst many researchers use The Cell to organize their own…
Read MoreWhere Did It All Go Wrong?! Quality Control For Your NGS Data
…If this number is high in your library it can indicate over-amplification or poor library-prep. (3) MiSeq QC: While not strictly a tool to check data from your own genome…
Read MoreWhere Did It All Go Wrong?! Quality Control For Your NGS Data
…If this number is high in your library it can indicate over-amplification or poor library-prep. (3) MiSeq QC: While not strictly a tool to check data from your own genome…
Read MoreSolid Phase Reversible Immobilization: How To Get A Bead On The Clean-up Of Your NGS Libraries
…concerned with reproducible library preps, gel purification after adaptor ligation is the step that requires the most attention to detail (see James Hadfield’s excellent instructions of what you need to…
Read MoreSize Analysis of High-Molecular-Weight DNA for Long-Read Sequencing
…essential to ensure the integrity of the starting sample by assessing DNA quality, shearing profiles, and library size. One way to do this is to analyze HMW DNA sequencing libraries…
Read MoreThe Next Big Thing: Alternative Polyadenylation
…extra steps, like end-repair, A-tailing and adaptor ligation steps. After synthesis of the second strand, cDNAs are selected by size and amplified by PCR and the resulting library is ready…
Read MoreRNAseq 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…
Read MoreA Colorful Route to Sequencing Success
…Collibri PS DNA Library Prep Kit for Illumina systems provide visual feedback that allows real-time tracking of the successful mixing of reagents at each step in the library preparation and…
Read MoreGenerating RNA-seq Libraries from RNA
…starts with size selection for small RNAs, ligation of selected fragments to adapters, and then generation of cDNA.4 New RNA Library Preparation Method A new RNA library prep method creates…
Read MoreCan’t Find pFavorite? Organize Your Plasmid Library
We’ve all been there. Digging through the -80°C freezer, fingers about to get frostbite as you scrape the ice off a tube to read an illegible plasmid name, hoping it’s…
Read MoreStudying the Epigenome by Next Generation Sequencing
…Thus, all that will remain is a library of fragments of methylated DNA, ready to be sequenced. Methyl-CpG binding domain protein sequencing (MBD-seq) The MBD family of proteins contains a…
Read MoreAn 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…
Read MoreChoosing a Scripting Language for Next Generation Sequencing: Python, Perl, and More
…difficult to read and understand someone else’s scripts. One benefit of Perl’s staying power is that the BioPerl library is more mature and larger than the BioPython library. Chances are…
Read MoreDNA 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…
Read MoreProbability Theory and Molecular Barcodes
…As an example, this approach enables parallel sequencing of different samples through the labeling of each library with a distinguishing barcode, or index (e.g. Illumina indexing). The procedure is straightforward:…
Read MoreZymo Research’s Mix & Go! Competent Cells: They’re Shocking… But Only to You!
…yield making them ideal for general cloning and library construction. TG1 and C600 strains are recombination competent. HB101 cells are excellent for library construction. 4. Choose your format: single aliquots,…
Read MoreBest Practices for DNA Shearing for NGS
Construction of high-quality sequencing libraries is pivotal to successful NGS, and DNA quality is one of the most critical aspects of library preparation. As this Nature Methods paper illustrates, DNA…
Read MoreNext Generation Sequencing: The Three Main Technologies
…Genome Sequencing (WGS), exome and amplicon studies. But first- some basics Almost all whole genome or exome sequencing is accomplished by first producing a fragment library, similar to shotgun cloning…
Read MoreSAGE Part 2: LongSAGE, RL-SAGE and SuperSAGE
…take 5 ?g of mRNA to create a library of hundreds of cDNA tags. In comparison, LongSAGE published in 2002 in Nature Biotech, could use 20 ?g of mRNA to…
Read MoreIgnore The Ticking Bomb At Your Peril: RNA-Seq Normalization- A Lurking Problem And It’s Solution.
You have spent days, if not weeks, at the bench setting up the treatment and control samples for that crucial experiment. You submitted your cDNA library for sequencing and after…
Read MoreWhy Is It Important To Run Your NGS Gels Consistently?
…low-input samples. Many NGS library preparation protocols use agarose gels for size selection of genomic DNA, RNA or final libraries. Gel electrophoresis is a simple size selection technique and is…
Read MoreBattle of the Methods: Whole Transcriptome Versus mRNA-seq
…extracellular RNAs — and even ribosomal RNA — that do not. But, ribosomal RNA is often depleted before library construction. Whole transcriptome analysis can take you from single base level…
Read MoreUsing PubCrawler: How to Speed Up Your Literature Searches by Crawling
…can also be a significant source of stress. In Search of a Library Assistant When I was a graduate student, I would diligently search PubMed for the newest articles published…
Read MoreNail Your Unmixing for Full Spectrum Flow Cytometry: 7 Top Tips
…record them into a reference library for reuse. DO: validate how long the reference library controls can be reused. The rule of thumb is one month, but you should validate….
Read MoreWhy DNA Size Selection Matters in NGS Pipelines
…improve assemblies, and even allow sequencing of low-input samples. Figure 1. Example of a well-sized DNA Library. Short-read sequencers, such as Illumina and Ion Torrent, operate best when fed DNA…
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