...Xiuhua Meng, Xiaolei Wei, and Ling Lu. (2016). “Highly efficient CRISPR mutagenesis by microhomology-mediated...

...requestors use the plasmid in their own labs. We highly encourage depositors to provide plasmid maps whenever...

...spectrophotometer uses the absorbance/transmission of light through a liquid to determine the concentration ...liquid. Molecules absorb different wavelengths of light to varying degrees and most have a specific wavelength... absorbance, you need to know the wavelength of light that your substance maximally absorbs. In the case...

...constructs, ADAR2DD(E488Q/T375G) (REPAIRv2) had the highest on-target efficiency and the lowest amount of off-target...

...vectors all have a genomic capacity that is much higher than other viral vectors (tens of kilobases; compared...

...with one or more of these parameters can lead to a high number of false negatives in Y2H screens. Testing...

...bacteria deploy proteosomes to degrade what the cell might consider junk protein. Though there are a number...

...some classic protocols on Protocol Online that you might find useful for your research. LabGuru Website: ...

...hosts. Also, self-transmissible plasmids may have high copy numbers and reduced metabolic load (Meyer 2009...

...amplification in the host cell. This enables relatively high and long-term expression of the reprogramming factors...

...externally. Fruit flies are also relatively straightforward to manipulate genetically. Scientists can make...

...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ...platform. Pour off the water in the sink. Take a brightfield image of the gel with an appropriate imaging ...25 kDa corresponding to the heavy chain (HC) and light chain (LC) proteins, respectively. There should ...

...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ...through a 0.2 µm PES filter. Aliquot and freeze upright at -20 °C Procedure Section 1: Affinity chromatography...and gently pour off the ethanol storage buffer. Tightly attach LabMate PD10 Buffer Reservoirs to the top...

...transduce multiple tissues. So how do you choose the right serotype? Keep asking questions!  Table 1: AAV Serotypes...

...bases. This is an example of a trace file from a high-quality portion of a sequencing reaction: Sequence...file from a portion of a sequencing reaction with high background: If you obtain a different sequence than...

...Distilled water Disinfectant Thermometer Water bath weights and floats Reagents None needed Procedure Ensure...be used to secure tubes in place and water bath weights can hold bottles in place. After putting your tubes...

...extinguisher Guidelines BSL-1 Guidelines Before You Work Right after entering the lab, put on the proper personal.... BSL-2 centrifugation steps require an aerosol-tight lid and rotors should be loaded and unloaded in ...

...reaction, incubation time can range from 45 mins to overnight. For diagnostic digests, 1-2 hours is often sufficient...and can happen for a variety of reasons, including high glycerol concentration. Learn more at (Link opens...

...Incubate at room temperature for 2hr, or at 16°C overnight (following the manufacturer’s instructions). Note...For many ligation reactions, especially if using "high concentration" ligase, 5min at room temperature ...

...your plasmid and order primers (see figure to the right). When designing your plasmid, think about what ...fragments can subsequently anneal to each other. Phusion High-Fidelity DNA Polymerase - incorporates nucleotides...

...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ... 0.2 µm PES syringe filter. Aliquot and freeze upright at -20 °C. Procedure Section 1: Seeding cells The...

...viral particles. Users may need to run lower or higher dilutions depending on their particular sample....appropriate PCR tubes. Pipette back and forth 5 times. Lightly cap the PCR tubes. Generate the Droplets Bring ...

...C for 1 hour, you can try incubating at -80°C overnight. Centrifuge for 20 minutes at 10,000 x g at 4°...

...well enough to be snug to your hand, but not so tight that you cannot perform your duties. Be sure to ...

...#3. Incubate plate with newly plated bacteria overnight (12-18 hours) at 37 °C. Pro-Tip Some plasmids ...

... 6 cm diameter LB/agar plates without antibiotic High concentration (100 mg/mL, 1000x) carbenicillin stock...

...Addgene's online resources. Career Videos Get career insight and advice from real Addgenies! Protocols (Link...

...also be used for PCR-mediated DNA mutagenesis. A higher cation concentration increases the error rate of...