...DNA elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you...protocols that recommend eluting in water, Tris-EDTA (TE), just Tris buffer, or some other variations. Does... affect your downstream experiment. TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer...the stability of your preparation for a long time. Tris buffer controls the pH, while the EDTA chelates ... enzymatic reactions, a valid alternative can be Tris buffer (with no EDTA) or water. These are two of... DNA through their kit. The advantage of using a Tris buffer is that it allows better control of the pH...for a short period, and there are indications that Tris buffer is better than water in these conditions....

...When cloning by restriction digest and ligation, you use restriction enzymes to cut open a plasmid (backbone... of how to do perform restriction digests.   Overview of the restriction cloning process. Both ...backbone will contain a variety of restriction enzyme cut sites (restriction sites) downstream of the promoter...DNA (insert) that has been cut by compatible restriction enzymes. An enzyme, DNA ligase, then covalently...sequence of interest (green, insert) are cut with restriction enzymes to generate compatible overhangs that...backbone covalent.  Before beginning the restriction digest and ligation process, you should carefully...these both must have compatible cut sites for restriction enzymes that allow your insert to be placed into...

...Thirteen: Guest Contributors of 2023 8 Great Depositor Research Stories Interested in contributing to the Addgene...bloggers and subject matter experts who have contributed to our blog in 2024. Thank you to…   Marty LaFleur...Kits We are very grateful for each and every contributor to the blog. Thank you! More resources on the...

...chromosome and for stringently controlled plasmids. Binding of IHF (green) redistributes DnaA (red) from ...chromosomal DNA.  Plasmids are said to be under stringent control when they depend on the host cell’s initiation...consider the replication mechanism of plasmids under stringent control, such as pSC101. DNA replication and regulation...appropriate time—for example, the cell must have enough nutrients available to complete the entire round of replication...assists with the process: it bends the DNA to redistribute the DnaA from high-affinity boxes to low-affinity...replication. A variety of other proteins also contribute to this regulation in different ways: methylating...DNA replication and regulation Plasmids must also strike a balance: replicating too slowly will eventually...

...email [email protected] if you would like to contribute to the catalog. The Catalog Will Start With Neuroscience...antibodies available at launch were created by James Trimmer, PhD, of the University of California Davis School... Figure 1: Antibody against PSD-95 from the Trimmer Lab. Information about the antibody, the target...Nguyen K, van der List D, Murray KD, Engebrecht J, Trimmer JS (2019) A toolbox of IgG subclass-switched recombinant...

...you to all who have contributed to the Addgene blog this year! Your contributions have helped researchers...: Erik Lee SnappSharing is Caring, But How to Distribute Open Hardware? Author: Benedict Diederich Course-Based...

...Have you ever tried digesting with XbaI or ClaI restriction enzymes and gotten unusual or unexpected ...methylation may affect your restriction digests. Why methylate? Restriction enoducleases are so ubiquitous...It turns out that restriction enzymes are one half of naturally occurring restriction modification systems... from the restriction modification systems, yet can still affect whether certain restriction enzymes can...associated with any restriction modification systems, their sequences may overlap with restriction sites, inhibiting...sequences to prevent them from being degraded by restriction endonucleases. A given prokaryote typically has... has genes encoding one or a few restriction modification systems containing methyltransferases that add...

...code, find and fix security-critical bugs, and contribute to constant updates of the software. In hardware...community to regularly replicate designs worldwide. Distributing hardware via globally established makerspace...

... Ingredient Concentration or % Purpose Tris HCl 62.5 mM Buffer SDS 2% (w/v) Detergent...buffer such as PBS or TBST. Stringent stripping The stringent stripping solution uses heat and detergent...Biosystems offer pre-made stripping solutions or kits. Mild stripping The mild stripping solution uses low pH...lose by trying to strip and reprobe. In this case, we recommend using mild stripping solution (see below...with the antibodies, especially with more stringent stripping methods. If you are planning to detect multiple...later, strip it thoroughly first. Then continue on with your western blot as normal! Stripping your membrane... to strip blots, and the best choice will depend on your specific experiment. Remember, stripping your...

...from the bioelectricity field In addition to sharing tools to study bioelectricity, bioelectric researchers...Developmental Bioelectricity symposium’s “What is Bioelectricity?” Journal of Bioelectricity ...of biology known as bioelectricity is revealing the important role of bioelectrical signaling mediated ...first introduced to bioelectricity when I attended the Developmental Bioelectricity satellite symposium... these slower-changing bioelectric states in living cells. Since bioelectrical signals are controlled ...Michael Levin. "The body electric 2.0: recent advances in developmental bioelectricity for regenerative and...  All living cells with a membrane possess an electrical state. This state, known as the resting potential...

... technologies distributed through Addgene over our first 10 years. 10 most distributed plasmid technologies...technology, but pSV-PGC1 was Addgene's first distributed plasmid and was created in the lab of my thesis...Bruce Spiegelman. It was also the only plasmid distributed in our 1st year, 2004, and it still gets requested...Plasmids Shipped! - By 2010, Addgene had hit our stride. We shipped our 100,000th plasmid. A variety of...up with all the new plasmids. So far, we have distributed over 10,000 CRISPR/Cas9 plasmids to more than...

...Researchers in developing countries often find themselves on the front lines of emerging global challenges...Seeding Labs to better serve those in developing countries. Developing an international scientific community...community Seeding Labs, a nonprofit founded in 2007, strives to expand the opportunities and scientific resources...resources available to researchers in developing countries--because scientific talent is everywhere, but ...around the world. As part of our own mission, we strive to help build the international scientific community... are always taking steps to increase plasmid distribution and disseminate knowledge around the globe. .... Addgene’s progress in international distribution As you can see from the chart below, we are making ...

...of how to analyze a restriction digest:     Restriction digest tips and tricks: The following tips ...plasmids. Here, we'll cover restriction digest analysis. Diagnostic restriction digest Diagnostic digests...Diagnostic restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction enzymes...to confirm the sequence of all the plasmids we distribute. This method is time-intensive, so we recommend...organization of different features within the plasmid. Restriction analysis can also be used successfully even if...relative sizes of the resulting DNA fragments. Restriction digests are commonly used to confirm the presence...backbone. To do this, you'll use enzymes with restriction sites that flank the insert. You will need to...

...providing plasmids to researchers in developing countries. We are proud to introduce the 5 scientists who...molecular biotechnology for improving expression of nutritionally and therapeutically significant proteins. In...Encoded H2O2 Probe with Improved Performance for Ratiometric and Fluorescence Lifetime Imaging” ACS Chemical...Expression Systems Learn about Global Plasmid Distribution ...

...alteration) Expensive buffers Tris-tricine 2.5–40 kDa Tris-tricine 30 V, 1 hour or 100 V, 1–2 hours...Limited protein size range Tris-acetate 40–500 kDa Tris-tricine 150 V, 1–3 hours Good for ...Running conditions Pros Cons Tris-glycine 6–400 kDa Tris-glycine 100 V, 1–2 hours Easy and...that Bis and Tris gels run proteins in different patterns, due to their differing chemistries. You therefore...-life; can alter proteins due to high pH Bis-tris 6–400 kDa MES for proteins <40 kDa 180 V, ... protein alteration) Expensive buffers Bis-tris 6–400 kDa MOPS for proteins > 30 kDA 200 V...therefore cannot compare Bis and Tris gels or blots to each other. Once you know your gel type, you’ll need...

...project We will ship you  2 µg of DNA in 20 µl of Tris buffer suitable for molecular biology experiments...this period, we’d love to hear from you if you’ve tried out this service! Let us know what you think. You...excited to begin offering this service to further contribute to our mission to “accelerate research and discovery...

...complex and dynamic field. We’re going to focus on a tried and true method of finding enhancers by isolating...mono-methylated and acetylated histones, but not tri-methylated histones, functions as an enhancer. We... me1, me3, and ac – denoting mono-methylation, tri-methylation, and aceylation, respectively (these ...suppose you have a piece string that’s 10 meters long. You twist and roll the string into a tight ball. Now...This post was contributed by guest blogger, Aneesh Karve, CTO at Quilt Data. This post was originally...follows: (mono_methylation + acetylation) - tri_methylation In the next section we’ll apply the above formula...Now the two ends of the string, instead of being 10 meters apart, are mere millimeters apart. Similarly...

... an exciting and emotional time!” George Church tries to recruit people who are nice as top criterion ...latest results as soon as the gels ran.” Dr. Ellis tries to lead his team as a scientists and a friend. George...

...end up stressed and rushing, no matter how hard I tried to build in extra time before the experiment. What...any experiment. That, more than anything else I tried, made a huge difference in my ability to focus, ...used Roberson Lab, UPenn. Western blotting. Retrieved Feb. 7, 2022. https://www.med.upenn.edu/robertsonlab...