...CRISPR 101 Series Learn about the PAM Requirements of Different Cas9 variants Learn about CRISPR Pooled.... & Liu, D. R. Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors....

...much trial and error to learn the scripting language, but our path forward was clear: to build one process...assembly algorithms meant that we already had a clear idea of the necessary steps to perform:    ...

...for Titering Your Lentivirus Learn All About Viral Vector Elements Learn How to Do Genome-Wide Screens... target cells. Poor tissue culture practices can lead to low titers and poor transductions. Cell lines...

..., using up valuable resources in the cell thus leading to slowed growth. Additionally, some protein products...well for most experiments, the lac promoter can “leak,” meaning that a low level of expression exists ...your favorite protein expression tips and tricks! Leave a comment below or browse Addgene’s collection of...

...a first time AAV user Learn about how AAV is used in retinal gene therapy Learn about the parts of an ..., would help improve split vector strategies and lead to greater and more consistent transgene expression...

...code inserted into the plasmid. This produces the least amount of variation between antibodies, allows for... induced to express antibodies. Antibodies are released into the growth medium and can either be used ...fragments - Fab fragments are created by enzymatically cleaving the variable regions of an antibody (each Fab ...they should be stored at -20 °C in aliquots of at least 10 µL. Antibodies cannot tolerate freeze-thaw cycles...single cells. Includes: Flow cytometry Read on to learn more about the applications common to each category...should be run with your protein of interest and at least one ubiquitously expressed protein such as GAPDH...protein:DNA complexes. After capture, the DNA is released from the complex and can be sequenced and amplified...

...very high titers (Zingg et al., 2017), which can lead to cells in far away brain regions expressing your...experiment. Some inducible constructs can exhibit leaky expression in the absence of the inducing drug, ... result in overexpression of your gene and could lead to toxicity or trouble interpreting your results...

...temperature 5-10 degrees above the primer and be at least 25 bp away from each primer, ideally located close...suitable for your cell lines and/or tissue types.  Please do note my oh-so-subtle use of reference genes,...can be used up quickly by abundant transcripts, leading to falsely lowered levels for less abundant transcripts...

...episode of our Hot Plasmids podcast series, you'll learn about optogenetics tools for controlling protein...plasmids covered in this segment Hot Plasmid 1: Photocleaveable PhoCl protein for controlling molecular processes...

...me: Drag-and-drop genome insertion, without DNA cleavage, with CRISPR-directed integrases🎵 On the 6th ...Addgene gave to me: dCas9-based gene editing for cleavage-free genomic knock-in of long sequences🎵On the...

...Guide to Synthetic Biology One of the best ways to learn about a new field is simply to know where to look...have additional resources that might be helpful, please reach out to me by email.  About the author Niko...

...this series, we’ll highlight one tip that we’ve learned in the lab that will hopefully make your life a...be able to locate materials quickly and easily. Learn more about using barcodes. Check out our “Lab...

...’ll discuss the types of programs SITN runs and learn how they think about communicating science. Listen... our Interview with Science Rapper Tom McFadden Learn about SITN's work at DayCon 2015 5 Reasons to Use...

...all of the answers, but we’re betting that your colleagues do. That’s why we’re launching the AAV Data Hub...thank you for your contribution to open science.  Please note that the AAV Data Hub is a beta project, and...

...really incredible science - I hope you've enjoyed learning about it (and maybe even picked up a new technique...a research story you want to share on our blog, please reach out to [email protected] - we'd love to hear...

...cleverness, and crafting skills have surpassed your colleagues. Success is sweet – as sweet as all that Halloween...yourself on in the coming days. Hopefully, I’ve made clear how excited we get for Halloween and how seriously...

...VSV-G, however, may be used across both systems. Please see our Biosafety page for information on working...enter the nucleus during mitotic breakdown of the nuclear envelope. Lentiviruses can transduce both dividing... are small enough to enter the nucleus through nuclear pore complex. This feature may be useful for the...interchangeably. For more information about lentivirus, please read our lentivirus guide. Are γ-retroviruses replication...stimulates the expression of transgenes via increased nuclear export. LTR Subcomponents: U3 R U5 in cis LTR; ...

...the next 100 years, many significant discoveries lead to the conclusions that genes encode proteins and...cell division, and each daughter cell receives at least one copy of the plasmid. By the 1970s the combined... tools in the molecular biologist’s toolbox. To learn more about different types of cloning methods check... ( ori ), an antibiotic-resistance gene, and at least one unique restriction enzyme recognition site. ...CRISPR technology. CRISPR is composed of a DNA endonuclease and guide RNAs that target specific locations...lentiviral particles. Visit our viral service page to learn more. Regardless of type, plasmids are generally...

...popular retroviral vector pBABE-puro. Read on to learn more about the fascinating history behind this vector...minimizing the chance of recombination that would lead to replication competent retrovirus.  When we arrived...

...Founders - Melina Fan, Ken Fan, and Benjie Chen. We learn what it was like to get Addgene off the ground and...idea got off the ground Chapter 3 - Addgene as a leader in standardized tech transfer and the adoption ...