We narrowed to 233 results for: pET

...agent (0.4 mM IPTG is a good start if you’re using a pET vector) and antibiotic for plasmid selection. Remove... for protein overexpression in E. coli, such as pET30a. If working with DNA that is refractory to enzymatic...your resultant plasmid libraries into chemically competent E. coli and analyze transformants for appropriately...order to reduce the chances of some clones being outcompeted by others in liquid culture, dilute and plate...transformation should be performed using a highly competent strain that you can use for long-term library ...probing for cell-binding activity.  First, cut out 20 Petri dish-sized circles of nitrocellulose membrane and...inhibitor cocktail. Line the bottom of 20 empty Petri dishes with thick filter paper and saturate each...

...available! Interested in purifying Cas9? Check out pET-28b-Cas9-His from Alex Schier's lab, designed for...generated mix of fluorescent proteins, determined by competing recombination events at the genetic level. Upon...

... These are sandwich, competitive, direct, and indirect. Sandwich and competitive ELISA use antibodies ...et al., 2019.   Competitive ELISA Similar to a sandwich ELISA, the competitive ELISA is also used ... be adapted to a competitive format. Figure 2: Overview of using Competitive ELISA to quantitatively...epitopes on the target protein, so they do not compete with each other. This antibody is also conjugated...to your protein of interest) will bind to the competing antigen in the wells while the protein-antigen...protein of interest in the original sample. The competitive ELISA is best used for detecting antigens that...sandwich ELISA. While the most common type of competitive ELISA is a modification of the sandwich ELISA...

...antibody can be used. In a competitive ELISA a multiwell plate is coated with a competing antigen (green).  The...antibodies are used.  Competitive ELISAs Scientists typically use a competitive or inhibition ELISA when... of ELISA are direct, indirect, sandwich and competitive. In a direct ELISA a reporter enzyme conjugated...sample, the less antibody available to bind to the competing antigen. In direct, indirect and sandwich ELISA... level of antigen in the sample whereas in a competitive ELISA the reporter signal is inversely proportional...sandwich ELISA. In this method, the target antigen competes with a reference antigen for binding to a primary... direct, indirect, and sandwich ELISAs, in a competitive ELISA the signal intensity is inversely proportional...

...scientific art competitions where creations can go on display for all to enjoy. The largest competition of this... University’s Annual Art in Science competition. This competition has gained so much public interest for...moonlighting as artists can compete in the Nikon Small World Photomicrography Competition - a 40 year old international...been featured in Time magazine . Other smaller competitions are now taking place and gathering momentum,...international competition that last year received over 2,000 entries. Do you mix your passion for art and...

...compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial...monomeric Venus pCEP4YPet-MAMM Yellow Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial...Bacterial Expresses CyPet with C-terminal His tag SCFP3A Cyan Mammalian Express a gene of interest fused...Bacterial Expresses YPet with C-terminal His tag SYFP2 Yellow Mammalian Express a gene of interest fused...

...genetic tools. Mobile-CRISPRi from the labs of Jason Peters, Oren Rosenberg, and Carol Gross provides a streamlined...Mobile-CRISPRi's modular components. Image from Peters et al., 2019 with permission.   Two methods... with an integrated copy of CRISPRi. Image from Peters et al., 2019 with permission.    The team showed...strains and environmental isolates. For example, Peters and colleagues used Mobile-CRISPRi to knockdown...possible with Mobile-CRISPRi. As a proof of principle, Peters and colleagues constructed a 40-member library ...microbiology 4.2 (2019): 212. PubMed PMID: 30675034. Peters, Jason M., et al. "Enabling genetic analysis of...PMID: 30617347. PubMed Central PMCID: PMC6424567. Peters, Joseph E. "Tn7." Mobile DNA III. American Society...

... Figure 1: Schematic of native (nuclease-competent) Cas9 interacting with either a full-length gRNA...relies on two distinct Cas9 variants—nuclease-competent or nuclease-null—in order to effect different ... to Cas9. For instance, when native (nuclease-competent) Cas9 interacts with a ≤15nt gRNA that contains... Figure 2: Gene activation with nuclease-competent Cas9. An RNA hairpin incorporated into the truncated...  Figure 3: Gene repression with nuclease-competent Cas9. When complexed with a truncated gRNA, Cas9...Cas9 is still able to bind DNA but is no longer competent to cut DNA. When directed to a site downstream...delivered into systems that already express nuclease-competent Cas9, obviating the need to generate new cell ...

...all bacteria are naturally competent to take up DNA, they can be made competent through chemical manipulation...Addgene’s blog to learn about making your own competent cells and our protocols page to learn about bacterial... The phagemids are packaged into replication-incompetent phage particles with assistance from a ‘helper... A., and Karen Guillemin. "Multiple phases of competence occur during the Helicobacter pylori growth cycle...Meibom, Karin L., et al. "Chitin induces natural competence in Vibrio cholerae." Science 310.5755 (2005):...

... and tested in the “market” by competing in business plan competitions. After much learning, the focus...plan competition and advance investors to the next stage of diligence." In business competitions everything...concept development and participating in business competitions. When the team was unable to secure enough money...CV tips - LabSpaces Blog Find a Buisness Plan Competition Good Relations - PR for the Biotech Business ...

...200 ul pipettor is set to 200 ul, wiped down with 70% ethanol, and a box of sterile truncated pipet tips...the pipettor, placed into wells, and partially dislodged from the agarose plug by rapid pipetting, as ...rapidly punch out cell clones with sterile truncated pipet tips. Indeed, an experienced operator can pick 48.... We highly recommend the use of an electronic pipettor with variable spacing (such as the Viaflo from...

...Prior to March 2020, internships were a fiercely competitive endeavor. Due to the COVID-19 pandemic, students...college, and graduate students. In order to gain a competitive advantage this cycle, consider the advice below...counterintuitive since that approach seems to create competition for your own spot. True! But you create a network...sense approaches that will prepare you for the competitive landscape beyond college. They will also serve...graduation. While these suggestions increase your competitive edge, they alone, are no guarantee of success...

...plasmids, they are generally electroporated into electrocompetent cells to ensure the highest possible transformation...use for your transformation(s). The amount of competent cells you need total and per transformation may...liquid culture, all of the bacteria can grow and compete in the same container, making it easier for a smaller...be its own colony, and will not be in direct competition with other colonies containing smaller, recombined...recommend ensuring the cells used are meant for repetitive sequences (such as STBL4 or Endura Duo), growing...

...curve, avoid pipetting small volumes (< 2 µL) or volumes too large for a standard micropipette (> 1,000 µL...2. When preparing a dilution series, use a new pipette tip for each step and mix the samples well by vortexing...readout. When preparing a standard curve, change pipette tips and mix thoroughly by inversion or vortexing...series, use consistent dilution factors, change pipette tips regularly, and mix samples very well.  Use...

...the same replicon are incompatible because they compete for the same replication control machinery within... concept is a little more complex than simply competing for replication control factors and tends to be...plasmids with the same centromere-like binding region compete for the same CBPs and NTPase to correctly partition...blocking replication of another and one plasmid outcompeting the other plasmid for replication or partitioning...machinery (Novick, 1987). In cases where one plasmid outcompetes the other, usually a smaller, higher copy plasmid...

...Sanger sequencing to confirm an insert that is repetitive or otherwise hard to sequence using NGS or long-read...such as G-C rich areas, secondary structures, or repetitive regions. Short-read sequencing can also be time-consuming...sequencing or Sanger sequencing, either because of repetitive regions or for higher accuracy of a particular...This method is great for resolving complex and repetitive sequences, but can be costly, low-throughput,...Nanopore sequencing to QC plasmids that are highly repetitive, such as those containing cassettes of the same...

...want to use electro-competent cells instead of the more common chemically-competent cells. You should perform...Follow the manufacturer’s instructions for your competent cells. For most standard cloning, you can transform...transform 1-2μl of your ligation reaction into competent cells such as DH5alpha or TOP10. If using much ...getting colonies, you might want to use higher competency cells. Additionally, if your final product is...

... appreciated the creative uses of old posters, pipette tip boxes, and more (stay tuned for a few blog ...sustainability in the lab!). The @ladiesinlabcoats used a pipette tip box and microcentrifuge tubes to make Eppi ...labchristmas #cute #reindeer #DeckTheLab #eppendorf #themuppets #scifun #funny #ChristmasScene #womenwhoscience...all who participated in the 2018 #DeckTheLab competition. The four winners will receive a holiday care...

...forming a vector that could create replication competent virus. Even though this system was efficient at...potential risk for the generation of replication competent lentiviruses (RCL) especially if you are working...positive cells could make the virus replication competent. The 3rd generation of lentiviral vectors was...8 kb) Potential for generation of replication-competent lentivirus (RCL) Efficient gene transfer Potential...Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human tropism can still infect...

...lab. Iron Chef-style competitions Every few months, we’ll have a cooking competition featuring a selected..., which is either an indication of the stiff competition or says something about my cooking.   This is... to organize. We’ll dedicate a post to these competitions in the future, and hopefully include some of...