Technical Resources

So you just used CRISPR Cas9 to edit a gene - how do you know you achieved the correct edits? The edits produced by using the CRISPR Cas9 system will not be identical and will not necessarily occur on all the alleles of a gene or cells in a population. In this video, we will introduce you to popular screening and validation strategies to determine if and which cell line has the correct edits. We will cover methods such as the mismatch cleavage assay, Sanger sequencing, and Next Generation sequencing. Our video will also take you through a basic screening and validation workflow that you can apply to your own gene editing project. This includes the basics of how to screen your polyclonal pool and how to isolate and screen monoclones. For more information on this topic, please visit: ➜ Knowledge Base: https://info.abmgood.com/crispr-cas9-screening-validation Want more tips, tricks, guides, and protocols on how to knockout a gene using CRISPR? Join our free, 4-week #CRISPRCrashCourse: ➜ http://info.abmgood.com/crispr-crash-course For a list of theoretical and practical CRISPR resources: ➜ https://info.abmgood.com/crispr-cas9 Connect with us on our social media pages to stay up to date with the latest scientific discoveries: ➜ Facebook: https://goo.gl/hc9KrG ➜ Twitter: https://goo.gl/gGGtT9 ➜ LinkedIn: https://goo.gl/kSmbht ➜ Google+: https://goo.gl/5bRNwC

So you've packaged and harvested your lentiviruses and you're ready to infect your target cells! But, what lentivirus transduction protocol should you use? Depending on whether you're infecting adherent or suspension cells or difficult-to-transduce cell types, we have different infection recommendations for you! In this video, we'll explain: → How to perform a lentiviral transduction experiment → An overview of the spinoculation and reverse transduction methods → How to confirm transgene expression Lentiviruses are extremely useful gene delivery systems for delivering cDNA, shRNA, and sgRNA to stably express, knockdown, or knockout your gene of interest in your target cells. Once you learn how to use lentiviruses for your own experiment, you'll be able to target not only dividing cells but also post-mitotic, non-dividing cells like neurons. → For more information about lentiviruses, access our free learning resources: https://info.abmgood.com/lentivirus-system → Access our full protocol for lentivirus transduction: https://www.abmgood.com/lentiviral-system-documents Request a free sample of our ViralEntry™ Transduction Enhancer: → https://www.abmgood.com/Transduction-Enhancers.html Watch our video "Multiplicity of Infection (MOI): What is it and how do I calculate it?": → https://www.youtube.com/watch?v=rfw_rcP5XOA Connect with us on our social media pages for fun educational materials introducing both practical and theoretical concepts in cell biology/life sciences: ➜ Facebook: https://www.facebook.com/appliedbiological.materials/ ➜ Twitter: https://twitter.com/abm_good ➜ LinkedIn: https://www.linkedin.com/company/applied-biological-material ➜ Instagram: https://www.instagram.com/abm_good ➜ Blog: https://info.abmgood.com/blog

Today we will be exploring the cell culture technique, Transfection. In this video, we will help you learn what transfection is, its application, ways to perform transfection, and go over a quick guideline for your consideration when preparing for your experiments! Here are some helpful time stamps for our Transfection video: 0:28 What is Transfection? 1:31 Applications 2:54 Consideration - What do you want to do? 3:29 Consideration - Knowing your cells to determine experimental conditions 4:31 Consideration - Plasmid Design 5:11 Consideration - Type of Transfection 6:14 Thank You! Connect with us on our social media pages for fun educational materials introducing both practical and theoretical concepts in cell biology/life sciences: ➜ Facebook: https://www.facebook.com/appliedbiological.materials/ ➜ Twitter: https://twitter.com/abm_good ➜ LinkedIn: https://www.linkedin.com/company/applied-biological-material ➜ Instagram: https://www.instagram.com/abm_good ➜ Blog: https://info.abmgood.com/blog

You're in the middle of preparing your cell cultures for an experiment but wait a minute - is that contamination? If so, who's responsible? Cell culture contamination is a pain but if it's not dealt with it can become an even bigger headache! In this video, we'll help you recognize the different calling cards left behind by the most notorious cell culture criminals: bacterial contamination, fungal and yeast contamination, mycoplasma contamination, and cell line cross-contamination. If contamination in your cell cultures is caught early, you might still be able to clean up the mess left by the culprits and save your precious cell cultures. ➜ Read more about how to detect and eliminate cell culture contamination in our "Cell Culture Quality Control and Contamination" article: https://info.abmgood.com/cell-culture-quality-control-contamination. Request your free sample of: • Mycoplasma PCR Detection Kit (Cat. No. G238): https://www.abmgood.com/pcr-mycoplasma-detection-kit-g238.html • Mycoplasma Elimination Cocktail (Cat. No. G398): https://www.abmgood.com/mycoplasma-elimination-cocktail-g398.html Learn more about our Cell Line Authentication Service: https://www.abmgood.com/Custom-Cell-Authentication-Service.html Connect with us on our social media pages for fun educational materials introducing both practical and theoretical concepts in cell biology/life sciences: ➜ Facebook: https://www.facebook.com/appliedbiological.materials/ ➜ Twitter: https://twitter.com/abm_good ➜ LinkedIn: https://www.linkedin.com/company/applied-biological-material ➜ Instagram: https://www.instagram.com/abm_good ➜ Blog: https://info.abmgood.com/blog

Don't let your cells die before the end of your project! In this video, learn how to develop an immortalized cell line from primary cells. Read our blog post for more information on cell line immortalization: ➜ https://info.abmgood.com/cell-immortalization Timestamps for topics covered in this video: [0:29] What are immortalized cells? [1:19] How do you generate immortalized cells? [3:47] Cell line quality control considerations You’re probably used to using cells that are taken directly from living tissue, called primary cells. The difficulty with primary cells is that their telomeres shorten after every cell division, causing the cells to enter senescence and stop dividing after only a few cell cycles. This means that if you are working on a long term project, you’ll frequently need to keep harvesting and re-establishing new batches of primary cells. In addition, every batch of cells is different due to different harvesting conditions, making reproducibility a headache! Immortalized cells (also called continuous cells or cell lines) are primary cells whose telomeres and/or tumour suppressor genes have been altered. Tumour suppressor genes (e.g. p53 and Rb) are important for signalling the cell to stop dividing when the likelihood of DNA damage is higher (i.e. after multiple cell cycles). In the case of immortalized cells, these genes have been knocked down or their function inhibited so that the cell is able to keep dividing indefinitely. In this video, we will cover two major cell immortalization strategies: (1) Telomerase Reverse Transcriptase protein (TERT) expression, and (2) viral oncogenes. We will also introduce you to two immortalization methods (lentiviral transduction and plasmid transfection). Connect with us on our social media pages to stay up to date with the latest scientific discoveries: ➜ Facebook: https://www.facebook.com/appliedbiological.materials/ ➜ Twitter: https://twitter.com/abm_good ➜ LinkedIn: https://www.linkedin.com/company/applied-biological-material ➜ Instagram: https://www.instagram.com/abm_good