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protocol icon Addgene Protocols


On this page you'll find practical lab protocols that you can use for a wide range of applications, with videos for select protocols in the right-hand column. You can find much more video content on the Addgene Video Page. We hope that you'll find these protocols useful in your own work. Please feel free to email us at [email protected] with any questions.

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Basic Molecular Biology

Name Description Link to Video
Making LB Agar Plates Create plates to culture bacteria in the lab Watch the Video!
Over-Agar Antibiotic Plating Quickly add antibiotic to a pre-poured plate Watch the Video!
Streaking Bacteria Isolate single bacterial colonies on an agar plate Watch the Video!
Inoculating a Liquid Bacterial Culture Prepare and grow bacteria in liquid medium Watch the Video!
Creating Bacterial Glycerol Stocks Store bacterial strains or plasmids for long term use
DNA Purification Miniprep, phenol-chloroform extract, and precipitate DNA
DNA Quantification Measure DNA concentration with a spectrophotometer
Restriction Digests Cut and analyze plasmid DNA with restriction enzymes
Agarose Gel Electrophoresis Separate DNA by size on an agarose gel Watch the Video!
DNA Ligation Assemble DNA using DNA ligase
Bacterial Transformation Introduce DNA into a bacterial strain Watch the Video!
Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Watch the Video!
Polymerase Chain Reaction (PCR) Basic PCR protocol with tips and FAQ
Purifying DNA from an Agarose gel Isolate specific DNA bands cut from a gel
How to Design a Primer Key considerations when designing primers
Sequence Analysis Verify important plasmid features using sequence analysis


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Plasmid Cloning

Name Description Link to Video
Restriction Cloning Assemble plasmids by restriction digest and ligation
Cloning by PCR Generate restriction sites by PCR
Modification by Annealed Oligo Cloning Add a short stretch of DNA to a plasmid
Gibson Assembly Combine overlapping DNA fragments in a single reaction
Ligation Independent Cloning (LIC) Scarless cloning with Type II restriction enzymes and T4 polymerase


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Virus

Name Description Link to Video
General Transfection Introduce plasmid DNA to mammalian cells
Lentivirus Production Produce lentivirus with a polyethyenimine (PEI) transfection protocol
Fluorescence Titering Assay for Lentivirus Quantify virus using fluorescence measurements
Colony Formation Titering Assay for Lentivirus Quantify virus by counting antibiotic resistant colonies
Generating Stable Cell Lines with Lentivirus Genomically integrate your DNA sequence into a cell line
Isolating a Monoclonal Cell Population by Limiting Dilution Generate monoclonal cell lines from a polyclonal pool of stable cells
AAV Production in HEK293 Cells Produce adeno-associated virus to deliver your gene of to the target of your choice
AAV Purification by Iodixanol Gradient Ultracentrifugation Purify adeno-associated virus from a preparation produced in mammalian cells Watch the Video!
AAV Titration by qPCR Using SYBR Green Technology Use qPCR to measure the physical titer of your adeno-associated virus preparation