We narrowed to 402 results for: PREP

...Resources on Addgene.org Find ready-to-use AAV Preps Find AAV plasmids Browse all viral vectors ...intracellular transport machinery. Addgene offers viral preparations of AAV2-retro vectors with a variety of transgene...

...Link opens in a new window) . Figure 1: Selection, prep, and testing of AAV enhancers in mice. Enhancers...

...cloning, and titering and testing your viral preparations. ... Video! Inoculating a Liquid Bacterial Culture Prepare and grow bacteria in liquid medium Watch the Video...long term use Watch the Video! DNA Purification Miniprep, phenol-chloroform extract, and precipitate DNA...Protocols for titering and testing your virus preparations. Name Description Link to Video General Transfection...Ultracentrifugation Purify adeno-associated virus from a preparation produced in mammalian cells Watch the Video! ...physical titer of your adeno-associated virus preparation Watch the Video! AAV Titration by ddPCR Use ddPCR...physical titer of your adeno-associated virus preparation Lentivirus Titration by ddPCR Use ddPCR to measure...

...able to start an overnight culture for plasmid DNA prep the following day. Tips and FAQ The optimal concentration...glycerol stock before you begin your plasmid mini-prep. Try not to freeze/thaw your glycerol stock too ...15-25% glycerol. You can prepare the glycerol stock the same time you prepare your plasmid DNA. In the...

...the prep with full (genome containing) particles. On average, iodixanol-purified AAV preparations contain...ultracentrifugation to get pure adeno-associated virus (AAV) preps for use in your research. Protocols...Ultracentrifuge T70i rotor or equivalent Reagents Opti-Prep (60% Iodixanol) QuickSeal tubes QuickSeal tube spacers...separate out contaminants from an impure AAV preparation. The 15% iodixanol step has 1M NaCl to destabilize...iodixanol solution is available under the name OptiPrep (Link opens in a new window) from Sigma-Aldrich...Centrifugal filter units (MWCO 100 kDa) Reagent Preparation 1 M NaCl/PBS-MK buffer Dissolve 5.84 g of NaCl... and 2 mL of 5 M NaCl to 43 mL PBS Procedure Preparation and loading of the iodixanol gradient: 15% iodixanol...

...polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously filtered) Microcentrifuge...conical tubes, VWR 21008-216 Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM...Protocol that can be used to titer lentiviral preparations carrying a fluorescence marker. Protocols...This protocol can be used to titer lentiviral preparations that carry a fluorescence marker. Determining... 75,000 cells into each well of a 6-well dish. Prepare a batch of cells as follows: Dilute 525,000 cells...thaw the lentiviral aliquot on ice prior to use. Prepare dilutions of the lentivirus into DMEM complete ...

...polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously filtered) Microcentrifuge...conical tubes, VWR 21008-216 Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM...This protocol can be used to titer lentiviral preparations that confer antibiotic resistance. Protocols...This protocol can be used to titer lentivirus preparations that confer antibiotic resistance. Determining...Determining the titer of your lentiviral preparation allows you to control the multiplicity of infection (MOI... Use this dose for the colony formation assay. Prepare a batch of DMEM complete containing 10 μg/mL polybrene...thaw the lentiviral aliquot on ice prior to use. Prepare 10-fold serial dilutions of the lentivirus into...

...NanoDrop to measure your samples, place 1-2µL of mini-prepped DNA onto the pedestal. Close the lid and click ...

...polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously filtered) Microcentrifuge...15 mL conical tubes, VWR 21008-216 Lentivirus preparation Appropriate antibiotic for selection (e.g. puromycin...puromycin, blasticidin) Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine (...determine the lowest dose that kills all of the cells. Prepare a batch of DMEM complete + 10 µg/mL polybrene by...Thaw the lentiviral aliquot on ice prior to use. Prepare a range of dilutions of the lentivirus in DMEM ...cell solution in this step. To seed the cells: Prepare a batch of cells as follows: Dilute 350,000 cells...growing well and have been sufficiently expanded, prepare cell stocks and/or harvest to test for protein ...

...you can find a protocol for kit-free plasmid mini-prep at the bottom of this page. Last Update: Feb. 8,...culture of bacteria . Pro-Tip Refer to appropriate DNA prep protocol for volume of bacteria to grow (low copy... Protocol: Kit-free Alkaline Lysis Plasmid Miniprep Prepare the following solutions: Solution I - Resuspension...pellet the bacteria before proceeding with DNA preparation. Pro-Tip If your entire overnight culture cannot...

...appropriate imaging system. Figure 1 Recombinant antibody preps should have 2 clear bands at ~50 kDa and ~25 kDa... Pro-Tip Addgene only uses recombinant antibody preps that have a purity of ≥90% but some labs use unpurified...prestained protein ladder on ice. Reagent Preparation Prepare IgG Standard Dilute the 2.5 mg/mL IgG isotype.... Add 8 µL of PBS to a total volume of 13 µL . Prepare the recombinant antibody Dilute or concentrate ...microfuge tube. Add 8 µL of PBS to total 13 µL . Prepare 1X MOPS buffer Dilute 25 mL of 20X MOPS buffer ...

...conjugating them in the lab? How will the sample be prepped, and will this or other factors affect epitope ...isotype. Overview of application Fix your samples. Prepare your samples using permeabilization, antigen retrieval...and controls follow that of IF (above). Sample preparation, signaling excitation, and assay readout vary...

... number of genome-containing particles in an AAV prep using qPCR, SYBR Green Technology, and Addgene’s... number of genome-containing particles of an AAV prep using SYBR green technology. This protocol was validated...each step Reagent Preparation Master Mix: Count the number of samples (n) and prepare master mix for an...μL Nuclease Free Water 4.7 μL 470 μL Procedure Prepare a plasmid stock of 2 x 10 9 molecules/μL to generate...

...plate. Be sure to pick multiple colonies for mini-prepping and verify insert by sequencing. Reference Page...

...unpublished materials. What about preprints? Addgene supports preprints and encourages authors to share ...alongside a journal article, a preprint, or without any paper at all. Posting preprints and openly sharing materials...reproducibility, and accelerate science. We encourage preprint authors to deposit materials without a hold or...or to release the hold as soon as the preprint is posted in order to support these goals. And we can update...article version any time later. We recognize that a preprint isn’t necessarily a clear sign that you are ready...your plasmids, so we won’t immediately release preprint-associated materials the way we will with journal...to consider sharing them openly as soon as that preprint is online!   Thanks for reading, and for depositing...

...confirm the identity and serotype of our AAV preparations. We recently published a description of the ... For labs that produce a small number of AAV preparations this level of quality control is sufficient ...facilities producing a large number of viral preparations in parallel, there is a very real risk of mixing...AAVs and the TA-based ligation and tagmentation preparation approaches described previously require a preliminary...effective substrates for tagmentation-based library preparation procedures. This finding eliminates the double-stranding...simplified approach, we sequenced over 100 AAV vector preparations and showed that VGS can quickly and reliably... script that can confirm the serotype of AAV preparations.  Using viral genome sequencing to determine...

...first scientific poster, but unsure how best to prepare. Maybe you’re a presentation veteran, but have ...experience. No matter the reason for attending, your preparation and day-of game plan will ensure you have the...experience possible. Tips for poster session presenters Prepare for success You’ve put a great deal of thought ...it’s time to expertly show off your science.  Prepare the night before: To avoid rushing out the door...poster as a visual aid, not a crutch. Even the most prepared of us can find ourselves talking to our posters... part of a poster session, but with a little preparation and a few deep breaths the day-of, you’ll handle...Strategically traverse the sea of posters If you come prepared with a list of posters of interest, then you’ve...

...talk to the occasion Don’t just plan to reuse a prepared talk unless it perfectly fits the occasion.  I...figure out how to make it suspenseful. 4. Start preparing your talk early - like today Don’t wait until ...for poor quality slides with sufficient time to prepare (and it does take time). For important talks, ask...loves a great animation. 8. Predict questions, prepare answers Speaking pros always have a few extra slides...come up with clear and concise answers. This preparation will help you avoid giving only yes or no answers...their phones. Speaking longer just makes you seem unprepared or arrogant. A bit short is always better than...

...consider using a different antibody.  Prepare a positive control Prepare and label a sample that expresses...paraformaldehyde-fixed cell samples, or other preparations. Sequential IF For a sequential IF experiment... ICC validated antibodies from Addgene), or be prepared to run some extra controls and validate the antibody...expect the target to be. You can test this by preparing a sample without the primary antibody, or with...different antibodies, blocking, washing, and sample preparation steps, in order to ensure your antibodies are...

... Untergasser Lab Preparation of chemically competent cells (protocol) Preparation of electrocompetent...Research to make competent cells because cells prepared using this kit can be transformed without heat...of electrocompetent cells (protocol)  Cells prepared using the Z-competent TM buffer can be easily ...