...PITCh-gRNA binding sites. This PITCh plasmic is cotransfected with a plasmid carrying Cas9, the PITCh-gRNA, ...Guide Pages Find CRISPR Plasmids for Your Experiments Deposit Your New Plasmid Tools with Us  ...Integration into Target Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about... knock-ins, some labs have used NHEJ for whole plasmid integration; however, since NHEJ is error-prone...cloning. An overview of PITCh. The PITCh plasmid contains a GFP-Puro cassette flanked by 5' and .... The PITCh-gRNA creates two DSBs in the PITCh plasmid to release the cassette, and an insertion-locus-specific...locus-specific gRNA The gRNA should target near the last coding exon of your gene of interest. For ideal ...

...for an estimated 2% of global plastic generation. Less than 9% of plastics are ever recycled, so adopting...mail. Recycle plastic packaging Packaging is a huge contributor to the increase in plastic manufacturing...services don’t offer plastic recycling, specialized companies can recycle plastic-- you just have to find...other scientists save resources by sharing plasmids Find plasmids at Addgene ...recycling! With the popularity of single-use items, plastic is a substantial portion of lab waste. Despite ...manufacturing, accounting for 40% of total plastic usage (Geyer et al., 2017). While most single-stream or city ...Corning takes back and recycles a lot of their plastic packaging (from Corning®, Falcon®, or Axygen® product...

...multi-subunit complex employed by class 1 systems. A putative new class 2 nuclease, Cpf1 (CRISPR from Prevotella...endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227. Find plasmids from this publication...: Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Figure from Zetsche et al.,...., 2015. How was Cpf1 found and tested? Class 2 CRISPR systems, including the type II Cas9-based system...Francisella), was annotated in several genomes and is classified as a type V CRISPR system. Like Cas9, Cpf1 contains...Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence....1 (2016): 31-34. PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik...

...CRISPR mediated genome engineering. 10. Plasmids 101: Control Plasmids Need help thinking about the types ...Scientists eBook Download the Plasmids 101 3rd Ed. eBook Read the Most Recent Plasmids 101, CRISPR 101, and Career... two eBooks (Management for Scientists and the Plasmids 101 2nd ed, the 3rd edition has now been released...top of the many developments in science and the plasmids, technologies, and researchers behind them. As...most popular posts continue to be those from our Plasmids 101 and CRISPR 101 series. These posts aim to ... the background they need to get started using plasmids and the revolutionary CRISPR technology in their...and how it’s used in CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate...

...developed the plasmids, pPSU1 and pPSU2. When restriction digested with PstI or EcoRV, these plasmids generate...Penn State Ladder: Homemade Plasmid Prep Penn State Ladder: Commercial Plasmid Prep Kit Amount 1000 Lanes...Two plasmids that can be used to make inexpensive 100 bp or 1 kb DNA molecular weight ladders were recently...polyacrylamide gels. Get Tips on Verifying Your Plasmid Table 1: Comparing Typical Comericial Ladders...are significantly cheaper. When using a homemade plasmid prep protocol, it cost less than $10 to make 1000... 100 bp or the 1 kb ladder. Using a commercial plasmid prep kit increases the costs to $20-30 per ladder...prep protocol here and the pPSU1 and the pPSU2 plasmids are available from Addgene. Learn How to Analyze...

...background via P1 transduction. Plasmid Expressing lambda red genes from a plasmid allows for a mobile recombineering... flexibility to modify the E. coli chromosome, plasmid DNA or BAC DNA.  To use the lambda red recombineering...bacteria with integrated defective prophage 2) from a plasmid, 3) from mini-λ or 4) from the lambda red phage...phage pL promoter.  It is a good idea to choose plasmids that also express the repressors associated with...limit leaky expression of the Red system.  Using a plasmid to express the lambda recombineering system is ... do this is to express lambda red genes from a plasmid with a heat sensitive origin of replication.  Once... them at 42℃.  Mini-λ A hybrid between using a plasmid and stable integration of a defective prophage ...

... labs and hinders experiments is mycoplasma contamination. Mycoplasma contamination has been shown to ...routinely test for mycoplasma contamination. There are several commercially available mycoplasma detection kits...larger microbes such as yeast, fungi, or bacteria, mycoplasma can be extremely hard to detect with levels reaching...cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients and can...the cell line’s growth and behavior. Signs of a mycoplasma contamination can be subtle and may include reduced...positive controls. Routinely test your cell line for mycoplasma and discard it if the culture has become overly...after transfecting with the transfer and packaging plasmids. A benefit of the multiple harvest strategy is...

...loved helping to answer your plasmid and cloning questions through our Plasmids 101 series and by responding...Melina Fan Plasmids 101: The Promoter Region – Let's Go! By Kendall Morgan Plasmids 101: Mammalian...Joanne Kamens Plasmids 101: Antibiotic Resistance Genes By Marcy Patrick Plasmids 101: Yeast Vectors...FAQs Answered! By Caroline LaManna Plasmids 101: What is a plasmid? By Margo R. Monroe Best wishes...as seen through the lens of Addgene’s blog and plasmid repository. Our blog is just over a year old, and...detailed information about this new technology. Our Plasmids 101 series continues to grab scientists’ attention...Genome Editing By John Doench (Guest Blogger) Plasmids 101: Origin of Replication By Kendall Morgan ...

...Chromatin-related genes with plasmids available from Addgene The links below pull up lists of plasmids containing the...containing lists of plasmids from the indicated publications. You can request all of these plasmids directly from...consolidate every conceivable chromatin-related plasmid in a collection that grows on a daily basis, the... does not have it. That said, if you know of a plasmid that we definitely don’t have that you really want...study chromatin and want to start sharing your plasmids through Addgene, don’t hesitate to start the process...these pages to learn when Addgene receives new plasmids containing these genes. As with the PIs above,... In-exhaustive Chromatin-related genes with plasmids on Addgene.org AGO1 BRCA1 CREBBP CUL1 H2AFX...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Read the paper in Science Find the new SHERLOCK plasmids More precision with dual-nuclease Cas9-Cas9 ...Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit for genome editing in plants...post Read the paper in Science Find the Cas14 plasmids Anti-CRISPR proteins for optogenetic control...paper in Nature Methods Find the anti-CRISPR plasmids Tagging human transcription factors with GFP...White’s lab recently deposited a set of CRISPR plasmids to tag human transcription factors with GFP. As...