...Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Background Information Bacterial... not need to obtain more competent cells and retransform. Bacteria on an LB agar plate can be stored at... temperature, can be found on your plasmid's information page. The next day you will be able to start ...freezing (5-6 times). Make sure that you see one uniform solution, and there are no layers present. Be sure...

...Follow this protocol to perform RNA extraction without a kit....Water-saturated Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl alcohol (49:1) 75% Ethanol RNase-free...phenol, mix thoroughly by inversion. Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) and then shake vigorously...Nanodrop), agarose gel, or bioanalyzer. For more information on nucleic acid quantification, see our protocol...from the homogenized sample(s). Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) per 1 mL of TRIzol® used...

...four biosafety levels. This protocol provides information for both biosafety level 1 (BSL-1) and biosafety...to your hand, but not so tight that you cannot perform your duties. Be sure to try out different sizes... and is being shared outside of Addgene for informational purposes only. If you choose to reuse or repurpose... can be held responsible for your use of the information contained in or linked to from these web pages...

... tips Bacterial Transformations We walk you through a standard plasmid transformation protocol and offer...tricks to reduce transformation time and increase efficiency Bacterial Transformation Protocol Inoculating...

...four biosafety levels. This protocol provides information for both biosafety level 1 (BSL-1) and biosafety...clearly marked as “BSL-2.” The names and contact information of the laboratory manager should be clearly visible... and is being shared outside of Addgene for informational purposes only. If you choose to reuse or repurpose... can be held responsible for your use of the information contained in or linked to from these web pages...

... are multiple objective lenses. Each objective performs essentially the same task but has different magnifying...an objective, you will see several pieces of information, but the most important (for now) is the magnification..., any stains used in your sample, and sample information so that you or others can interpret your images...scratches, and dust can easily impair microscope performance. Be sure to clean up your work area and microscope...

...your purified virus. Nonetheless, we recommend performing a buffer exchange before using the purified AAV...filter and store at 4 ℃. 0.001% Pluronic-F68 (formulation buffer) Add 5 μL of 100X Pluronic F-68 to 49.95...not easily removed. After each spin, add more formulation buffer and sample and make sure to pipet back...is less than 500 µl, bring up the volume with formulation buffer. Use a P1000 to the bottom of the filter...

...Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary Oligo overlap cloning can be used anytime...that can be added when ordering them or can be performed enzymatically later. Experimental Procedure Digest...the vector with 0.75-6 ng of annealed oligos). Transform 2-3μL into your favorite competent bacteria and...

... the Video! DNA Purification Miniprep, phenol-chloroform extract, and precipitate DNA DNA Quantification...Ligation Assemble DNA using DNA ligase Bacterial Transformation Introduce DNA into a bacterial strain Watch... virus using fluorescence measurements Colony Formation Titering Assay for Lentivirus Quantify virus by...

...specific products or equipment. Inclusion of this information is solely for transparency intended to support... counter 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Vortex Stir bar...flask.Incubate in a 37 °C, 5% CO 2 incubator on a shaking platform set to 120 rpm. *Pro-Tip* Do not use cells that...

...specific products or equipment. Inclusion of this information is solely for transparency intended to support...spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Clamp stand and...divide the sample among multiple columns. For information about the total binding capacity for your columns...

... Gel Electrophoresis DNA Ligation Background Information Gel purification allows you to isolate and purify...step with the now gel purified DNA. Find more information about DNA quantification here . Tips and FAQ ...

...Protocol, tips, and FAQ for how to perform a restriction digest of plasmid DNA...more enzyme is advisable. Reactions are often performed with 0.2-0.5 µL of enzyme because it is difficult...

...containers Reagents Liquid for pipetting Background Information Anatomy of a Pipette This front view photo of...the Volume on the Pipette Although each pipette performs the same function, the numbers are read differently...

...Standard protocol for performing agarose gel electrophoresis, including tips to improve resolution and...Watch the protocol video below to learn how to perform gel electrophoresis. Equipment Casting tray Well...

...Follow this protocol to perform digital droplet PCR (ddPCR) for lentivirus vector titration....Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence Titration...

...Restriction Digest of Plasmid DNA Background Information During several different stages of molecular ...

... capabilities. Taking into consideration the information above, primers should generally have the following...

...also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture...

...Plasmid DNA from Bacterial Culture Background Information Plasmids can carry one or more antibiotic resistance...