We narrowed to 160 results for: ALC
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TypeBlog Post...two successful mutations in a single cell. We calculated the likelihood of obtaining homo- and heterozygous...
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Filming Science Videos in the Age of Social Distancing
TypeBlog Post...good once. Even when things return to relative normalcy, I’ll be incorporating that mobile monitor stand... -
Antibodies 101: Multiplex Immunofluorescence
TypeBlog Post... the different signals in a certain region or calculate a colocalization factor to assess the spatial ... -
Plasmids 101: Restriction Cloning
TypeBlog Post...base pairs for each varies, it is difficult to calculate this based on DNA concentration alone. One method... -
9 tips for a successful postdoctoral experience
TypeBlog Post... that all group leaders must master. While the calculus of these decisions is complex, here are a few ... -
Important Considerations When Using AAVs
TypeBlog Post...non-enveloped viruses, so they are resistant to alcohol-based disinfectants. A 10% bleach solution should... -
FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution
TypeBlog Post...Hartmann, and Peter Hegemann. "Rhodopsin-regulated calcium currents in Chlamydomonas." Nature 351.6326 (1991... -
Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein
TypeBlog Post...optimized to achieve high knock-in efficiency. To calculate the tagging efficiency in the transfected cells... -
Custom CRISPR Screens & the Green Listed Software
TypeBlog Post...The different labs use different algorithms to calculate which would be the best gRNAs. Interestingly, ... -
Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins
TypeBlog Post..., et al. "Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging." Nature... -
Antibodies 101: Introduction to Antibodies
TypeBlog Post...concentration of the known samples is used to calculate the concentration of protein in the experimental... -
Chemogenetics Guide
TypeGuide... phospholipase C, which releases intracellular calcium stores. There are currently three Gq DREADDs based... -
How to Deposit Your Plasmids with Addgene
TypeBlog Post... Lysine (D123K), lowers biosensor affinity for calcium". Please describe any tags or fusions (e.g. His... -
Deep Dive: qPCR
TypeBlog Post... Standard Curve? Amplification Efficacy Calculation Amplification Efficacy Assumptions Standard... -
Optogenetics + CRISPR, Using Light to Control Genome Editing
TypeBlog Post...light-inducible heterodimerizing cryptochrome 2 (CRY2) and calcium and integrin-binding protein 1 (CIB1) proteins.... -
Cre-lox system
TypeCollection...pLV-EF1-Cre-PGK-Puro Cre EF-1 alpha Lentiviral Alcudia 108639 fli1aCreERT2 Cre-ERT2; endothelial reporter... -
Neurodegeneration Plasmid Collection
TypeCollection...proteinopathy, ALS Nicolas Fawzi 98664 MBP-hnRNPA2_deltaLC HNRNPA2B1 His, MBP T7 Multisystem proteinopathy... -
Lentivirus ddPCR Titration
TypeProtocol... forget to factor this into your calculations later (see calculation example below). Seed 300,000 cells... a CSV file which will be used to calculate the titer. Calculations In the experimental setup above, the...20 200 40 400 80 800 160 1600 To calculate the titers, first calculate the number of viruses per cellular...glutaGRO, Corning 25-015-CI) 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...over copies\ RPP30}$$ Use the viruses/genome to calculate the infectious titer: $$T = {V*C*D\over v}$$ Where...titer obtained from the appropriate dilutions to calculate the final infectious titer. Tips and Troubleshooting...green). To increase the accuracy of the titer, calculate an average of several dilutions. Figure 1: ddPCR... -
CRISPR Library Amplification
TypeProtocol...new vented falcon tube for every two transformations. Each of the four 14 mL Vented Falcon Tubes should... mL of LB Agar per bioassay plate. 14 mL Vented Falcon Tubes (BD Biosciences, 352059) 3X LB Agar + Antibiotic...supports normal growth is expected to work. 50 mL Falcon Conical tubes (Fisher, 14-432-22) Reagent Preparation...Aliquot 3 mL SOC into each of four 14 mL Vented Falcon Tubes and have 1 mL SOC per electroporation readily...all liquid from cuvette and add to 14 mL vented Falcon Tube containing 3 mL SOC. Repeat for each of the...DNA-Endura from cuvette. Shake four 14 mL Vented Falcon Tubes at 30-37 ℃, 225 rpm for 1 hour. After the... -
Fluorescence Titering Assay
TypeProtocol... of PBS. Calculate the fraction of fluorescent-positive cells in each well. When calculating titer, only...mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...remaining well, a cell count is required for calculating the titer. Incubate for 48–72 h. Gently aspirate...leading to underestimation of the true titer. Calculate the transduction units per mL (TU/mL) using either... -
Isolating a Monoclonal Cell Population by Limiting Dilution
TypeProtocol...CI) Heat-inactivated FBS 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...will be used to seed the 96-well plate. Sample Calculation First, use a hemocytometer to quantitate the ...well plate, make 10 mL of a 5 cell/mL solution. Calculate the total cells needed: Total cells needed: 10... -
Kit Free RNA Extraction
TypeProtocol...Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl alcohol (49:1) 75% Ethanol RNase-free water or TE solution...by inversion. Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) and then shake vigorously by hand for 10...homogenized sample(s). Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) per 1 mL of TRIzol® used. Shake vigorously... -
AAV Production in HEK293 Cells
TypeProtocol...Thermo Fisher, 12605010) 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...incubator for 24 h–36 h. Proceed with transfection: Calculate the amount of each plasmid needed to have a 1:... Therefore, for each plasmid we need: Sample Calculations RepCap: 0.08 μg/bp × 7,265 bp = 582.1 μg Volume... -
Centrifugation
TypeProtocol... the two, use a conversion calculator or chart like the G Force calculator (Link opens in a new window... -
Protocol - How to Ligate Plasmid DNA
TypeProtocol.... Use a (Link opens in a new window) ligation calculator to easily quantify how much vector and insert...vector end. (Link opens in a new window) Ligation calculators are easily found on the web. Just enter the concentration... -
Colony Formation Titering Assay
TypeProtocol...mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...integration events decreases as the dilution increases. Calculate the transduction units per mL (TU/mL) as follows... -
AAV Titration by qPCR Using SYBR Green Technology
TypeProtocol...Addgene. The values highlighted below in red were calculated using this plasmid, but will change if you use...use a different plasmid. Sample Calculation Size in bp of Addgene plasmid #59462 : 6208 bp Concentration... -
Plasmid Cloning by PCR (with Protocols)
TypeProtocol...TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this calculator we get a final Reverse Primer sequence of 5’-...base pairs for each varies, it is difficult to calculate this based on DNA concentration alone. One method... -
Coomassie Purity Stain of Recombinant Antibodies
TypeProtocol...trendline for the standards. Google Sheets can calculate this for you as follows: In the Chart Editor ,...based on your sample’s HC + LC area. Example calculation: If the sample HC + LC Area is 18179.442 and ... -
DNA Quantification
TypeProtocol...Calculate your DNA sample concentration and purity using Addgene’s DNA Quantification Protocol....