...Dec;11(12):1261-6. doi: 10.1038/nmeth.3147. Bioluminescent pH sensor Understanding intracellular pH regulation...pRSETb-pHlash and pcDNA3.1+-pHlash, that use Bioluminescence Resonance Energy Transfer (BRET) were developed...simultaneously acquired from the the fluorescent and luminescent siganls and a ratio is calculated (the BRET ratio...voltage sensitivity and speed, and an endoplasmic reticulum export motif and trafficking sequence for improved...dynamics and interactions between the endoplasmic reticulum (ER), mitochondria, cytoskeleton, and endosomes...PMID: 26456817. Nano-lanterns for multicolor luminescence imaging  Fluorescent imaging techniques have...The Nano-lanterns rely on the principle of bioluminescence resonance energy transfer (BRET), whereby photons...

...the plasmid DNA from the column using a particular buffer or water. See column manufacturers for more detail...DNA; filter the remaining plasmid DNA through a column that binds the plasmid DNA and lets other materials... detail. Find columnless purification protocols on our website. 6. Verify the plasmid After purifying ...

...chosen focal volume and thus the microscope captures only fluorescence coming from this volume. The consequence...organism to make the complicated transition from a clump of cells to a fully differentiated body. In contrast...of dendritic cell extension into the small bowel lumen in response to epithelial cell TLR engagement." ...

...minor 475 nm peak, respectively. When wtGFP is illuminated with UV-violet light (~400 nm), the chromophore...PA-GFP to display almost no fluorescence when illuminated with 488nm light. However, when UV-violet light... to a 100-fold increase in fluorescence when illuminated with blue light (1). Similar to PA-GFP, scientists...derived from DsRed and mRFP, respectively. When illuminated with violet light, both variants are photoconverted...

.... Click on it, select your desired serotype and volume, and submit your request.    Figure 1: The...AAV2, AVV5, AAV8, or AAV Retrograde (AAVrg), and volumes of 1 mL, 2.5 mL, or 5 mL. Please note that all ...orders will be shipped in 100 µL aliquots, so the volume options will appear as 10, 25, or 50 units.  Once...

... the sequence of interest. 5. Use a silica spin column for purification of the samples you send for DNA...isopropanol precipitation method or using a silica spin column available from several vendors. The precipitation... I would strongly recommend using a silica spin column. They can be pricey, but well worth it. 6. Edit...

...descending order, in the case of numeric columns.  The data in any column are searchable and you can search ...You can access the datatable here. Current table columns are target gene, species, sequence, Addgene plasmid...

... sharing concerns. Ask alumni of the lab. Most labs have websites listing alumi and it is easy to use ...ignore warning signs If lab members, neighbors or alumni give you information about the PI that concerns...

...contributed by Theresa Liao of the University of British Columbia. When transitioning from an academic science career...Good luck! Resume 101 from University of British Columbia, Student Services - includes videos on general... of Physics & Astronomy, University of British Columbia. Originally on track for a research career, she...

...locally. Your university probably has an alumni directory. Alumni who enter their information in these searchable...information so you can tap them for advice. As a future alum, these are people you can “cold call” and ask for...

... complete plasmid sequencing using an in-house Illumina MiSeq. The raw sequencing reads are assembled ...particularly difficult to sequence and assemble using the Illumina MiSeq platform, such as GC-rich regions and highly...short 200–250 base pair reads sequenced on the Illumina MiSeq.  Resolving QC issues If the QC team identifies...

...plexWell technology to easily and quickly create Illumina sequencing libraries. From start to finish the...libraries are then QC’d, pooled, and prepared for Illumina sequencing on our newly donated MiSeq.  For Complete... also adds the necessary adapter sequences for Illumina sequencing as well as unique barcodes per sample...

...having to re-run the western.  Remember that chemilumescent methods, like horseradish peroxidase protein...membrane) Increase time of wash step Increase volume of wash buffer Incomplete stripping (If stripping...Underexposed Increase exposure time Chemilumenescence: use a more sensitive exposure kit Too...Overexposed blot Shorter exposure time Chemilumenescence: use a less sensitive exposure kit No ...

...saturation density, decreased rates of growth, or clumping. To ensure the best possible results for any tissue...approach is that it tends to result in a higher volume of lower titer virus. In such cases you may want...them! Changes in such variables as cell density, volume, incubation time, and MOI can all impact the success...

... full plasmid sequencing at our facility using Illumina MiSeq Next Generation Sequencing (NGS) technology...need to modernize our approach to handle larger volumes of data. In this two-part blog series, we will ...continuous updates. The generous donation of a Illumina MiSeq sequencer enabled us to perform the necessary...

...what if an FP fusion with a resident endoplasmic reticulum (ER) protein instead localizes to the cytoplasm...control of secretory proteins in the Endoplasmic Reticulum and live cell imaging approaches. Erik is also... inheritance independently of the endoplasmic reticulum." The Journal of cell biology 154.2 (2001): 317...

...barcoded, locus-specific libraries compatible with Illumina next generation sequencing in order to study evolutionary..., with each plasmid carrying a unique barcoded Illumina-M13F bridging primer. These bridging primers are... are amplified and used to generate Illumina sequencing libraries using a 2-step PCR-based protocol and...

...into the oviduct wall between the ampulla and infundibulum. In vivo electroporation is performed immediately...oviducts. After solution injection into the oviductal lumen, tweezer-type electrodes are placed to mediate electroporation...i-GONAD is that it requires a high-concentration, column-purified DNA template.   Many thanks to Channabasavaiah...

...mechanisms that generate luminescence. This can then be detected by a luminometer. Non-specific DNA stains...sensitivity but low specificity (Degeling, 2012). Bioluminescence assay kits are available from different biotech...

...and fluorescent light sources. Visible light illuminates each cell as it passes through and scatters in...  Tips for flow cytometry success Break up cell clumps Success in a flow cytometry experiment depends ... through a nylon mesh cell strainer to break up clumps before running on the instrument. To sustain cell...