...of 15% iodixanol step 5 mL of 25% iodixanol step 5 mL of 40% iodixanol step 3 mL of 60% iodixanol step ...and loading of the iodixanol gradient: 15% iodixanol step: mix 4.5 mL of 60% iodixanol and 13.5 mL of 1 ...buffer 25% iodixanol step: mix 5 mL of 60% iodixanol and 7 mL of 1X PBS-MK buffer and 30 μL of phenol red 40%... 40% iodixanol step: mix 6.7 mL of 60% iodixanol and 3.3 mL of 1X PBS-MK buffer 60% iodixanol step: mix... Purification by Iodixanol Gradient Ultracentrifugation AAV Purification by Iodixanol Gradient Ultracentrifugation...medicine, iodixanol can be used as an intravenous isomolar contrast agent. In research, iodixanol is used...On average, iodixanol-purified AAV preparations contain ~20% empty particles. 60% iodixanol solution is...

...qPCR Using SYBR Green Technology AAV Titration by qPCR Using SYBR Green Technology You may also like......particles in an AAV prep using qPCR, SYBR Green Technology, and Addgene’s AAV Titration protocol. Protocols...genome-containing particles of an AAV prep using SYBR green technology. This protocol was validated using an internal...

...-saturated phenol-chloroform Chloroform 100% ethanol or isopropanol 90% ethanol 70% ethanol TE buffer ...plasmid-containing supernatant. Add either ethanol or isopropanol to precipitate the plasmid DNA. Either ... Optional: Perform phenol-chloroform extraction - see protocol below . Note: Phenol-chloroform extraction...When phenol is mixed with the aqueous solution containing DNA, proteins will move into the phenol phase... either 700 μL of cold 100% ethanol or 350 μL room temperature isopropanol to the solution to precipitate...Protocol: Phenol-Chloroform Extraction of DNA Samples Add an equal volume of TE-saturated phenol-chloroform.... Note: Phenol-chloroform is a hazardous waste - DO NOT pour down sink. Top Protocol: Ethanol Precipitation...

...Water-saturated Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl alcohol (49:1) 75% Ethanol RNase-free water...). You can use either Isopropanol or Lithium Chloride for this step. Isopropanol (Option A): Add 1 volume...). You can use either Isopropanol or lithium Chloride for this step. Isopropanol (Option A): Add 1 volume...using an all-in-one acid guanidinium thiocyanate-phenol solution such as TRIzol®. For the precipitation...precipitation step, two options are also included: using Isopropanol or Lithium Chloride. This protocol was adapted...vol) N-laurosylsarcosine (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol® or similar product such as TRI Reagent...decontamination solution like RNase AWAY® or RNaseZAP® Isopropanol (for precipitation step, Option A) 7.5 M Lithium...

...15 min in 20% ethanol in deionized water. To prepare 20% ethanol, dilute 2 mL of ethanol into 8 mL of ...Microcentrifuge tubes BCA Assay, Thermo Fisher 23227 β-mercaptoethanol 4X protein loading buffer Precast SDS-PAGE ...-PAGE running buffer Prestained protein ladder Ethanol iBlot 2 PVDF Mini Stack, Thermo Fisher IB24002 ...sample for loading as follows: Add 10% v/v β-mercaptoethanol to the 4X protein loading buffer. Dilute 4X...

...the lab. Equipment Water Bath 10% bleach or 70% ethanol Distilled water Disinfectant Thermometer Water ...water. You may use a 10% bleach solution or a 70% ethanol solution to wipe down the inside of the water bath...marker that is also resistant to chemicals such as ethanol. Writing on the tops of the tubes and lids of items...

...produced in mammalian cells AAV Purification by Iodixanol Gradient Ultracentrifugation Protocol Mulitchannel...preparation. AAV Titration by qPCR Using SYBR Green Technology Protocol Agarose Gel Electrophoresis Gel electrophoresis... process when requesting samples from Addgene Technology Transfer How to Order A step-by-step walkthrough...

...use Watch the Video! DNA Purification Miniprep, phenol-chloroform extract, and precipitate DNA DNA Quantification...the target of your choice AAV Purification by Iodixanol Gradient Ultracentrifugation Purify adeno-associated...Video! AAV Titration by qPCR Using SYBR Green Technology Use qPCR to measure the physical titer of your...

...rProtein A Gravitrap columns and gently pour off the ethanol storage buffer. Tightly attach LabMate PD10 Buffer...of Protein A/G binding buffer and store in 20% ethanol at 4 °C . Pro-Tip Columns may be re-used up to ...

...Sterilize your lab bench by spraying it down with 70% ethanol and wiping it down with a paper towel. Maintain...

...appropriate materials. For example, you may use 70% ethanol, quaternary ammonium, or bleach solutions. Ask ...

... for all. Alternative fixation methods such as methanol or acetone may be better for some applications...

...working flame. Spray down the bench with a 70% ethanol solution and wipe down with a paper towel. Count...

...medium appears orange/yellow if using a standard phenol red pH indicator) because the buildup of waste ...

...Trypsin EDTA (TrypLE, Thermo Fisher, 12605010) Ethanol, VWR, EX0276-1 Benzonase 250 U/µl, Millipore #71205...

...gene delivery in vivo. Zufferey R et. al. Nat Biotechnol . 1997. 15(9):871-5. PubMed . Self-inactivating...