Search results for: library prep
The Ins and Outs of Protein Concentration – Chromatography
…protein at the end of the prep. Maltose binding protein (MBP), glutathione-s-transferase (GST), or histidine tags are the most useful for this purpose. The only other prerequisite for this method…
Read MoreCan You Stand the Cold? Cryosectioning for Beginners
Before you can perform histology on your tissue samples – you need to prep them. This means you must fix them, embed them, and section them into thin slices for…
Read MoreThe Nature of Denaturing (Protein Gels, that is!)
…frequently are gummy, particularly if the protein prep is from cell or tissue extracts and therefore contains DNA. Boiling homogenates your sample, as the heat melts any DNA in the…
Read MoreThe Key to Unlocking DNA from FFPE Tissues
Formalin fixed paraffin embedded (FFPE) tissues are valuable samples that typically come from human specimens collected for examination of the histology of biopsies for the detection of cancer. But each…
Read MoreKnow Your Western Blot Jargon! A Quick Review.
…a fraction of your protein. After extracting as much of your protein as possible you must prep them for electrophoresis. This is typically done by mixing them with sample/loading buffer…
Read MoreTop Ten Tips for TAP
…TEV cleavage, you may find that the prep contains TEV protease (28K, if His-tagged ~53K) and IgG heavy chain (~53K). This is not important if you want to stain the…
Read MoreTips and Tricks to Get Around Low Plasmid Yields in Agrobacterium tumefaciens
…for their kits if you make enquiries. Do bear in mind though, that even with kits, your yield may be quite low (5-20 ng/µl). Furthermore, and from experience, your prep…
Read MoreAAV Production Part I: Tips for Setting Up
…for 30 minutes. Spin in a microcentrifuge at 14,000 rpm, 4°C for 30 minutes. Remove and keep supernatant. This is your crude AAV prep. If you’re using a plasmid expressing…
Read MoreFree PCR for 5 Years (or How to Make your Own Taq and Pfu Polymerase)
…simple, prep can easily generate hundreds of thousands of units of highly purified polymerase per liter of culture. Put another way, one batch can provide a lab of 10-12 people…
Read MoreDIY Electrocompetent E. coli
…to make great DIY electrocompetent E. coli prep. In this article, you’ll discover a protocol for making DIY electrocompetent E. coli strains that you can use for electroporation. I’ve also…
Read MoreMolecular Docking: Let the Docking Do the Talking
…1. Protein or Receptor Structure Prep To start with, one needs to obtain a 3D-structure of the protein (Protein Databank Structures or homology models). Then you add the correct number…
Read MoreSix Steps to Successful Mouse Genotyping
…from the cells. Use a ‘dirty’ DNA prep for most mouse genotyping PCR protocols. They are quick, easy, and inexpensive. An example is the HotSHOT technique of DNA preparation: boil…
Read MoreTroublesome Site-Directed Mutagenesis: Troubleshooting Your Experiment for Stubborn Mutations
…PCR product you are using in the transformation. Plate several concentrations of your transformed suspension (e.g. 10 µl of bacterial prep, 20 µl, 50 µl, 100 µl) and only pick…
Read MoreHow to to Speed up Commercial Midi and Maxi Plasmid Preps
…prep time, using coffee filters… Yes, using coffee filters and funnels purchased from any grocery store, the precipitated genomic DNA, protein and cell debris is filtered out prior to applying…
Read MoreNGS Quality Control in RNA Sequencing- Some Free Tools
…an annotated reference genome. rRNA One key measurement is the amount of ribosomal RNA (rRNA). This is important since the removal of rRNA by various RNA sample prep protocols is…
Read MoreThe Art of Size Selection for Small RNAs
…comparing size selection methods for miRNA and small non-coding RNA discovery. The scientists, from McGill University and the European Molecular Biology Laboratory, compared the Pippin Prep from Sage Science to…
Read MoreAll in the Chip: Ion Torrent Sequencers
…PGM and Proton machines, library construction is like other sequencing processes, including DNA fragmentation, end polishing with enzymes, and ligation of adapters. After that, things get a little different. Amplifying…
Read MoreLight-up RNA Aptamers: Illuminating the World of RNA
…with SELEX New aptamers are obtained from “systematic evolution of ligands by exponential enrichment” (SELEX). In brief, SELEX is an extensive oligonucleotide library consisting of randomly generated sequences flanked by…
Read MoreAnalyzing RNA-Seq Data
…reads to a reference sequence, or known genome, will show similarities and differences. Strand specificity: Some library preparation approaches allow for the retention of strand-specific information so that aligned cDNA-derived…
Read MoreFree Online Bioinformatics Tools
…and software tools. It was created in 2006 by the Molecular Biology Information Service for the Health Sciences Library System at the University of Pittsburgh and can be accessed from…
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