We narrowed to 400 results for: POR C

...spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Clamp stand...Equipment Class II, Type A2 Biological Safety Cabinet 4 °C Refrigerator Pipette controller Benchtop centrifuge...phosphate dibasic (NaH 2 PO 4 ) Store up to 1 month at 4 °C Adjust pH to 7.0 Autoclave or filter sterilize Protein...µm PES filter. Aliquot and freeze upright at -20 °C Procedure Section 1: Affinity chromatography Equilibrate...A/G binding buffer and store in 20% ethanol at 4 °C . Pro-Tip Columns may be re-used up to 5x when purifying...446 Benzamide, Millipore Sigma 12072 Antipain, Millipore Sigma 10791 Leupeptin, Millipore Sigma L2884 Aprotinin...monobasic monohydrate, Millipore Sigma 71507-250G Sodium phosphate dibasic, Millipore Sigma S7907-500G 1 ...

...parameters: Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Enzyme Activation 95 10...titer: $$T = {V*C*D\over v}$$ Where: T = Infectious titer, TU/mL V = Viruses per genome C = # Cells per ...Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 Pierceable...Benzonase in DMEM complete to each well. Incubate at 37 °C for 30 min. Gently aspirate media from wells, using...can be used for ddPCR immediately or stored at -20 °C until ready to use. Preparing for ddPCR Thaw samples...EX0276-1 Benzonase 250 U/µl, Millipore #71205-3 Polybrene 10 mg/mL, Millipore, TR-1003-G Molecular Biology...place the PCR plate with the foil onto the metal support block. Place the block in the plate sealer and ...

...Michael, S., Kuleshova, E., Ojala, D., Huang, C. C., Gerfen, C. R., Schiller, J., Dudman, J. T., Hantman,...Technology References Burger, C., Gorbatyuk, O. S., Velardo, M. J., Peden, C. S., Williams, P., Zolotukhin... Ravi, N., Wu, W. L., Sánchez-Guardado, L., Lois, C., Mazmanian, S. K., Deverman, B. E., & Gradinaru, ...) Liu, D., Li, T., Liu, L., Che, X., Li, X., Liu, C., & Wu, G. (2024). Adeno-associated virus therapies... to display neuronal retrograde transport, AAVrg. Retrograde transport refers to the direction of movement...nuclear export. Poly(A) signal in cis The polyadenylation or poly(A) signal aids in the nuclear export of ...soma to axon) and retrograde (from axon to soma) transport. Many rAAV vectors have been engineered to express...

...attention. Let agarose solution cool down to about 50 °C (about when you can comfortably keep your hand on ... with a pipette tip. Place newly poured gel at 4 °C for 10-15 mins OR let sit at room temperature for ... set more quickly if you place the gel tray at 4 °C earlier so that it is already cold when the gel is...period of time; b) using a wider/thinner gel comb; or c) loading less DNA into the well. Another method for... be in the bottom portion of the gel, but all of the EtBr will be in the top portion and your bands will...overboil the solution, as some of the buffer will evaporate and thus alter the final percentage of agarose...

...individual tubes of 5, 10 or 20μL for storage at -20°C. Whenever you need to set up ligations in the future...optimization. Incubate at room temperature for 2hr, or at 16°C overnight (following the manufacturer’s instructions...efficiency of ligation can be improved by incubation at 37°C. Proceed with bacterial transformation . Tips and ...desired insert. If you are in this situation, it is important to treat the digested vector backbone with a phosphatase...setting up the ligation reaction itself, it is important to determine the amount of cut insert and vector...being sure to increase the amount of buffer proportionally. 1μL of ligase should be sufficient for larger...

...heat-inactivated FBS and 5mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and FBS lots can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...dose of antibiotic, which may not be stable at 37 °C. To achieve a stable cell pool, the antibiotic selection...well plate and left undisturbed for 13 days. (a, b, c) Colonies formed by expansion of single cells for ...CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...of cell death upon antibiotic selection. It is important to monitor these cells regularly and replace the... the surviving cells in the culture, so it is important to do regular media changes and maintain optimal...

.... M., Chen, I. S., Hahn, W. C., Sharp, P. A., Weinberg, R. A., & Novina, C. D. (2003). Lentivirus-delivered...PMID: 33768124 Marshall, H. M., Ronen, K., Berry, C., Llano, M., Sutherland, H., Saenz, D., Bickmore, ...PMID: 18092005 Merten, O., Hebben, M., & Bovolenta, C. (2016). Production of lentiviral vectors. Molecular...Schambach, A., Zychlinski, D., Ehrnstroem, B., & Baum, C. (2013). Biosafety features of lentiviral vectors....., Kochenderfer, J. N., Norberg, S. M., Hinrichs, C., Highfill, S. L., Somerville, R. P., Panch, S. R....transcription of the full-length virus after it is incorporated into a host genome. This is often found in second-generation...cells, they kill off any cells that have not incorporated the lentiviral genome and the cells that survive...

... Cepko, 2020). Incubate the mix for 1 hour at 50 °C or follow manufacturer's instructions. You can purchase...Gibson DG, Smith HO, Hutchison CA, Venter JC, Merryman C. (2010). Chemical synthesis of the mouse mitochondrial....1515 (Link opens in a new window) Rabe BA, Cepko C. (2020). A Simple Enhancement for Gibson Isothermal...other. Phusion High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed... product by restriction digest . Sequence the important regions of your final plasmid, particularly the...

...parameters. Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Denaturation 95 10 2 1 ...sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume (20 µL) C = Copies/µL D ...Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 48-well...place the PCR plate with the foil onto the metal support block. Place the block in the plate sealer and ... reading. When the droplet reading is complete, export the data from all wells as a CSV file and use the...

...Wong, G. K., Cameron, E. R., Kilbey, A., & Neil, J. C. (2016). Gamma-Retrovirus integration marks cell Type-Specific...10.1371/journal.pone.0154070 PMID: 27097319 LaFave, M. C., Varshney, G. K., Gildea, D. E., Wolfsberg, T. G....gkt1399 PMID: 24464997 Maetzig, T., Galla, M., Baum, C., & Schambach, A. (2011). Gammaretroviral vectors:...Rubat, L., Nikoniuk, A., Macmorland, W., Horlock, C., Matsumoto, S., Williams, S., Smith, K., Price, J...cells, they kill off any cells that have not incorporated the retroviral genome and the cells that survive...the only type of selectable marker. Fluorescent reporters, such as GFP, are another common selection marker...interest and contains the sequences that will be incorporated into the host cell genome but cannot produce...

...Whitehead, E. H., Guimaraes, C., Panning, B., Ploegh, H. L., Bassik, M. C., Qi, L. S., Kampmann, M., &...Delivery Validate Genetic Edit References CRISPR ( C lustered R egularly I nterspaced S hort P alindromic...25408407 Dixit, A., Parnas, O., Li, B., Chen, J., Fulco, C. P., Jerby-Arnon, L., Marjanovic, N. D., Dionne, D...testing multiple pegRNA designs is particularly important for prime editing. CRISPR Intereference : Target...Predicted On-Target and Off-Target Activity Another important factor to keep in mind when designing your gRNA...In addition to off-target activity, it is also important to consider factors that maximize cleavage of ...generally perform by chemical transfection or electroporation. Expression can be transient, or you can generate...

...stored at 4 °C for short-term storage. For long-term storage they should be stored at -20 °C in aliquots...conjugated to fluorophores should never be stored below 4 °C. Certain isotype subclasses, such as IgG3, aggregate... after thawing and therefore should be kept at 4 °C. Record and track lot numbers for all antibodies used...necessary in cases where extreme specificity is important. Their epitope specificity allows binding to a...antibodies only bind a single epitope, so it is important to choose a monoclonal antibody that will recognize...using a secondary against a tag, such as His, incorporated in the sdAb. sdAbs can have weak signals due...with conjugating reactions are possible. It is important to research your reaction and buffer before conjugating...

...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...CI) Heat-inactivated FBS Polybrene (10mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...

...media at 37 °C (for at least 15 minutes). Prewarm 3X LB Agar + Antibiotic plates at 37 °C. Prewarm 8X ...Chill a box of 200 µL micropipette tips in a -20 °C freezer. Aliquot 3 mL SOC into each of four 14 mL ... mix. Electroporate cells (one at a time for a total of eight electroporations): Electroporator Conditions...and have 1 mL SOC per electroporation readily available for post-electroporation recovery of cells. Ensure...if the recombinant band makes up a significant proportion of the DNA pool. Last Update: August 17, 2023...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents.../or increasing the skewness of the pool. 8 electroporation cuvettes (BioRad, Micropulser, 0.1 cm ) 20 ...

...Patron at The Sainsbury Laboratory Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. A modular cloning...10.1371/journal.pone.0016765. PMID: 21364738 . Engler C, Youles M, Gruetzner R, Ehnert TM, Werner S, Jones...Plasmids with control elements and fluorescent reporters to be used in combination with the MoClo-YTK toolkit...

...inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...

...standard Before Starting Warm the hot plate to 100 °C. Thaw IgG standard and prestained protein ladder on...microcentrifuge. Heat the samples for 10 min at 100 °C in a heat block. Spin the sample briefly in the microcentrifuge...IgG standards in Google Sheets. Open exported CSV file. Select import. Choose file. Format as needed. Select...that sharing the full details of our protocols supports reproducibility and accelerates science. Here,...information is solely for transparency intended to support reproducibility in science. Last Update: June 14...intensity of the protein bands to the overall lane. Import the gel image into ImageJ. Select File . Select... each peak by selecting the center of the peak. Export results as a csv file. In the results table, select...

...longer period of time; b) using a wider gel comb; or c) loading less DNA in the well. How do you get better...around the band as possible. To do so, it is often important to take the excised band, lay it down on the UV...sides with the razor blade. This is especially important during the DNA purification step, as many kits... fragment. The weight of the gel is directly proportional to its liquid volume and this is used to determine...manufacturer's instructions. Note: It is usually important to determine the concentration of the DNA that...

... refrigerators to achieve a temperature between 4°C and room temperature. This general protocol suits ...on the surfaces of the water bath as the water evaporates from the tub. Disinfectant may be added to prevent...cover on the top of the water bath to prevent evaporation and maintain the desired temperature. This also...

...software to ensure a melting temperature between 50-60°C for your PCR primers. Step 2: Linearize Vector In ...provided by your polymerase manufacturer. It is very important to remove all free nucleotides from your PCR product...