...ImageJ or similar software Reagents Purified recombinant antibody, 0.9–1.1 mg/mL SimplyBlue SafeStain,... bio-safe Coomassie G-250 stain (Addgene uses SimplyBlue SafeStain) and ImageJ software. The sample is...and reagents from other vendors should produce similar outcomes when using these protocols. However, please... Pour off the water in the sink. Add 20 mL of SimplyBlue SafeStain and incubate for 1 h with gentle agitation...agitation on a rocking platform. Pour off the SimplyBlue SafeStain in the sink. Add 100 mL of deionized...Analysis Use ImageJ (Link opens in a new window) or a similar photo software to determine the relative intensity...

...cloning sites, will result in similar sized bands, thus making this simple digest less informative. This...Plasmid Size -OR- Insert and Backbone Size The simplest form of diagnostic digest is one in which you ...products from each orientation are different sizes. Simply run the digest products on a gel and those clones...

...choice with minimal limitations. Background In its simplest form, PCR based cloning is about making a copy...TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but we need to use the reverse complement to get... get PCR amplification. We can start similarly, taking the final 18bases of the ORF, including the stop...primer. If you are amplifying from a plasmid or simple template, there is very little chance for mis-priming...

... (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol® or similar product such as TRI Reagent®, RNAzol®, QIAzol® ...how effective your cell lysis protocol is. While simple homogenization is effective for most mammalian ...Homogenize or lyse tissues or cells in TRIzol® or a similar product. For tissues: use 1 mL of TRIzol® per 100...

...bands needed to be separated - see FAQs below . Simply adjust the mass of agarose in a given volume to...How do you get better resolution of bands? A few simple ways to increase the resolution (crispness) of ...you get better separation of bands? If you have similarly sized bands that are running too close together...

...primers are mainly used for amplifying a small, simple fragment of DNA. On the other hand, a long primer...The structure of the primer should be relatively simple and contain no internal secondary structure to ...

...do you get better resolution of bands? A couple simple ways to increase the resolution (crispness) of ...you get better separation of bands? If you have similarly sized bands that are running too close together...

... Your Primers Primer design for LIC is often as simple as using the backbone manufacturer's suggested ...the first G on 3'-5' strand from cut site). For simplicity, only the 5' primer is shown here. Note: Use ...

...RNA extractions. Plus, centrifuges are useful for simply collecting liquid to the bottom of a tube. You ... can be about bench height. They often spin at similar speeds to their smaller cousins, but can hold much...

... al. , 2010). Number of fragments assembled simultaneously Multiple fragments can be assembled in one ...opens in a new window) Rabe BA, Cepko C. (2020). A Simple Enhancement for Gibson Isothermal Assembly. bioRxiv...

...can create thousands of identical copies using a simple set of reagents and a basic heating and cooling...Your 5’ and 3’ primers should be designed to have similar melting temperatures (Tm). Set the annealing temperature...

...and reagents from other vendors should produce similar outcomes when using these protocols. However, please...plate washer (optional) Scale Microsoft Excel or similar software Reagents Purified antigen Purified recombinant...

... to the BSL-1 requirements. Conclusion Although simple, following appropriate PPE protocol goes a long...

...tags or shRNAs to any vector (the procedure will simply differ in terms of primer design). Let's assume...

... trace file will give you more information than simply looking at the bases provided by your sequencing...

...pipette and avoid inconsistent pipetting with our simple tips Bacterial Transformations We walk you through...

...containers that can be autoclaved. Conclusion Although simple, following appropriate BSL-1 and BSL-2 protocol...

...sites . Sometimes enzymes cut sequences which are similar, but not identical, to their recognition sites....

...their own competent cells. This is a relatively simple procedure and is useful for performing low efficiency...

...the molarity of DNA ends available for ligation. Simply put, there are only two ends on any given piece...