...polymerase pLKO.1 - TRC Cloning Vector Cloning protocols for using the pLKO.1 vector, a backbone used by the ...addgene.org with any questions. Intro to the Lab Bench Introductory techniques designed to help you get started ...and Two (BSL-1 and BSL-2) Safety measures for laboratories operating at BSL-1 and BSL-2 Watch the Video...Video! Pipetting Learn about selecting the correct pipettor and pipette tip, how to dispense liquids, and ... its use Weighing Reagents Learn how to weigh laboratory materials on a balance Basic Molecular Biology...Watch the Video! Creating Bacterial Glycerol Stocks Store bacterial strains or plasmids for long term use ...

...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...marker. Determining the titer of your lentiviral vector allows you to control the multiplicity of infection...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Inverted...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tips Different brands and lots of FBS...protocol was developed using low titer lentiviral vectors (10 5 TU/mL). If you anticipate that your viral...units per mL (TU/mL) using either the dilution factors (method 1) or the volume of virus (method 2): Method...Fraction of cells with fluorescence D = Dilution Factor V T = Transduction Volume, mL For example: If 150,000...

...filter. Aliquot 50–100 µL and store at -20 °C. Aliquots can be thawed and stored at 4 °C prior to use. Thawed...into sterile tubes. Store the tubes at -80 °C. After thawing the solution can be stored at 4 °C for up to...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...

...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents...heat inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...Swirl gently to mix and place the dish in the incubator. Incubate for 48–72 h. Pro-Tip Different cells...colonies/total volume in the well (mL) x dilution factor e.g., If the 1:100,000 well has 75 colonies, then...the indicated serial dilutions of the lentiviral vector pRosetta . The following day, cells were treated...the indicated serial dilutions of the lentiviral vector pHAGE EF1α dCas9-KRAB . The following day, cells...

...volume. Below that is the tip ejector button which is used to push the tip ejector down the pipette to remove...Learn about selecting the correct pipettor and pipette tip, how to dispense liquids, and how to handle...liquid (think: 0.1 µL to 1 mL). When working in a laboratory, properly dispensing liquid ensures the accuracy...the set volume for the the pipette, and the tip ejector ring at the bottom which pushes the pipette tip...dedicated waste container and pressing on the tip ejector button. If pipetting biohazardous materials, be...

... plates) and store at -20 °C. Once a standard is thawed do not freeze it again but store at 4 °C and use...SYBR Green Technology You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Introduction...the titer of samples obtained from academic viral vector cores. Our titers were similar to those reported...30 min Equipment qPCR instrument Heating plate Pipettors 1–10 µL single channel pipette 20–200 µL single...uL) Volume of nuclease free water (uL) Dilution factor Total dilution Dilution 1 (DNase step) 5 uL AAV...

...amplification protocols that have been developed by the depositor for a specific pooled library available at Addgene...Addgene. We recommend following the depositor's specific protocol if one is available. If a pooled library...require modifications dictated by the originating laboratory for optimal results. If you obtained the pooled...amplification protocol is expected to obtain satisfactory results for libraries up to 200,000 individual...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents...time for a total of eight electroporations): Electroporator Conditions: Bio-Rad Micropulser Ec1 0.1 cm ...

...also adhered to. None of the authors, contributors, administrators, or anyone else associated with Addgene...and bathroom. These items should remain in the laboratory. When traveling between lab spaces, take your...been designed by the Addgene nonprofit plasmid repository and is being shared outside of Addgene for informational...

...Creating Bacterial Glycerol Stocks for Long-term Storage of Plasmids Recovering Plasmid DNA from Bacterial...temperature. Now screw on the top of the bottle and store the LB at room temperature. When ready to grow your...bacterial culture at 37°C for 12-18 hr in a shaking incubator. Note: Some plasmids or strains require growth... overnight incubation. Optional: For long term storage of the bacteria, you can proceed with Creating ...

... Protocols Video protocol guides for standard laboratory procedures. How-To Videos Screencapture walk-...enzymes Diagnostic Restriction Digest Protocol Storing and Handling Addgene Plasmids What to do after ...receive your plasmids from Addgene Instructions for Storing and Handling Plasmids Genomic Deletions with CRISPR...Electrophoresis Gel electrophoresis is the standard laboratory procedure for separating DNA by size for visualization...

... mM EDTA Store Solution I at 4°C Solution II - Denaturing Solution 0.2 N NaOH 1.0% SDS Store Solution ...acetate 23 mL glacial acetic acid 57 mL of dH 2 O Store Solution III at 4°C Grow 2 mL overnight cultures... room temperature before quantifying and using. Store DNA at 4°C. Tips and FAQ Plasmid purification kits...

...sites. Often, the size of the plasmid insert and vector backbone are known and thus this technique can ...Restriction Digest If you clone an insert into a vector by a single restriction enzyme, you will need to...which orientation the insert is ligated into the vector backbone and therefore the only thing left to do...

...cut the vector, you will need to use a phosphatase to prevent re-circularization of the vector if you ...having difficulty finding an enzyme that cuts your vector's multiple cloning site (MCS), but does not cut ...

...water Microscope slide Anti-fade mounting medium Laboratory wipes 15 mL conical tubes 50 mL conical tubes...paraformaldehyde and follow your institution's laboratory safety guidelines for disposing of waste in the...remove the coverslip. Blot the coverslip with a laboratory wipe to remove excess liquid. Add 1 drop of anti-fade...

...research and are found in many different types of laboratories. These tools allow you to observe specimens ... in the future. Conclusion Like any important laboratory instrument, you should be sure to take care of... up your work area and microscope after use and store the microscope appropriately....

...your plates are ready for use or storage at 4 ℃. You should not store your plates for longer than 1 month...material to reduce condensation. The plates are then stored at 4 ℃ until use. Once your plates have solidified...

...loop Bunsen burner (or other small flame source) Incubator Marker Reagents LB agar plate (with appropriate...resistance and your initials. Labeling within a laboratory setting is important for organization, and it...

...spreading of antibiotic makes it easy for an investigator to conveniently plate and select transformed...spreading Bunsen burner (or other small flame source) Incubator Reagents 6 cm diameter LB/agar plates without ...

... lab! Introduction A water bath is a piece of laboratory equipment that helps bring your materials to .... Water baths can also be placed in walk-in refrigerators to achieve a temperature between 4°C and room...

...a blank chart to your spreadsheet. In the Chart Editor , select the Chart Type drop down menu and select...calculate this for you as follows: In the Chart Editor , select the Customize tab. Select the Series drop...