...larger conical tubes. Depending on the centrifuge their speed ranges can vary, but are typically lower ... the two, use a conversion calculator or chart like the G Force calculator (Link opens in a new window...Tabletop Centrifuges sit on your benchtop and typically hold the common 1.5 mL microfuge tubes used in...

... Addgene Protocols On this page you'll find practical lab protocols that you can use for a wide range...Generating Stable Cell Lines with Lentivirus Genomically integrate your DNA sequence into a cell line ... SYBR Green Technology Use qPCR to measure the physical titer of your adeno-associated virus preparation...AAV Titration by ddPCR Use ddPCR to measure the physical titer of your adeno-associated virus preparation...Lentivirus Titration by ddPCR Use ddPCR to measure the physical titer of your lentivirus preparation Antibodies...

...depends on your application. A diagnostic digest typically involves ∼500 ng of DNA, while molecular cloning...largely determined by the volume of DNA to be cut. A typical restriction digestion reaction could look like ...reaction in 1 hour. Using this ratio, you can calculate the minimal amount of enzyme for your reaction...the vector if you are cloning in an insert. CIP (calf alkaline phosphatase) or SAP (shrimp alkaline phosphatase...enzymes cut sequences which are similar, but not identical, to their recognition sites. This is due to "...

...significantly and should be empirically determined for your sample. Typical ratios may range from 1:1 to...pipettes Benchtop centrifuge compatible with 50 mL conical tubes Automated cell counter 37 °C, 5% CO 2 incubator...antibody plasmid DNA 500 mL vented flask 50 mL conical tubes Microcentrifuge tubes Cell counting chamber.... Transfer the HEK293 cells and media to 50 mL conical tubes. Centrifuge for 15 min at 3100 x g to pellet...

...polymerase that can withstand radical temperature changes during a typical PCR. The DNA polymerase has ...template), a researcher can create thousands of identical copies using a simple set of reagents and a basic...GC-rich templates. What does each ingredient specifically do? Template DNA: Contains the portion of DNA...Buffer with MgCl 2 : Provides an optimal and stable chemical environment for the DNA polymerase to work adequately...

...stable cell lines Equipment Class II, Type A2 Biological Safety Cabinet 0.5–10 µL single channel pipette...mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...tissue culture treated dish, Corning 3516 15 mL conical tubes, VWR 21008-216 Lentivirus preparation Appropriate...You Start The health of the target cell line is critical for obtaining accurate results. Check the cells...and/or harvest to test for protein expression. Typically, cells transduced with lower dilutions of the ...

...make bacteria more susceptible to either chemical or electrical based transformation, generating what are...antibiotic containing agar plate. This step is not critical for Ampicillin resistance but is much more important...large plasmids (>10 kb) or BACs, what can I do? Chemically competent cells are fast and easy to use, but...

...should have identical sequences on the ends (sequences A and B in the figures). These identical sequences...PCR with primers that contain a 5′ end that is identical to an adjacent segment and a 3′ end that anneals..., Hutchison CA, Venter JC, Merryman C. (2010). Chemical synthesis of the mouse mitochondrial genome. Nature...

...visualization and purification. Electrophoresis uses an electrical field to move the negatively charged DNA through...eye protection and gloves when working with this chemical. Note: If you add EtBr to your gel, you will also...approximately 75-80% of the way down the gel. A typical run time is about 1-1.5 hours, depending on the...polyacrylamide gel electrophoresis (PAGE), which is typically used to separate 5 - 500 bp fragments. How do ...

...medium. This dot is composed of millions of genetically identical bacteria that arose from a single bacterium...draw in discontinuous lines. Start by streaking a vertical line of bacteria along one edge of the plate. ...

...polymerase can create overhangs of varying length (typically 10-12 bp) based on a specific sequence. The annealed... the replication cycle. Empty vectors for LIC typically employ type II restriction enzymes (e.g. BsaI)...back the guanosine residue and become stalled. A typical T4 Pol reaction is shown. Mix the components (...

...lab! Introduction Microscopes are emblematic of biological research and are found in many different types...suggests, light microscopes take advantage of the physical properties of light to detect small objects. Two...the light follows from the source to your eyes is called the light path . Light travels from the light source...

...antibiotic-containing agar. This protocol will focus specifically on selection with carbenicillin; however it ...required for effective over-agar selection has been empirically determined. See selection curve below. Carbenicillin...

...microfuge tube. Using a scale, weigh the tube with the gel fragment after zeroing the scale with an empty tube...

...materials to a particular temperature, catalyze chemical reactions such as restriction digests needed in...sure to use a marker that is also resistant to chemicals such as ethanol. Writing on the tops of the tubes...

...culture Aseptic Technique Aseptic technique is a critical skill for success in numerous fields such as microbiology... AAV Titration by qPCR Use qPCR to measure the physical titer of your adeno-associated virus preparation...

...mounting medium Laboratory wipes 15 mL conical tubes 50 mL conical tubes Before Starting Refer to the manufacturer's...

... using, you will be rotating dials behind the numerical display or the top of the plunger of the pipette... to pipette, it’s important for scientists to calibrate their pipettes to make sure that they are properly...

...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 ...

...the pellet. Air-dry the pellet for 5-10 minutes. Critical It is important to not let the pellet get too ...