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.

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 | Watch the Video! |
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 | Watch the Video! |
Purifying DNA from an Agarose gel | Isolate specific DNA bands cut from a gel | Watch the Video! |
How to Design a Primer | Key considerations when designing primers | Watch the Video! |
Sequence Analysis | Verify important plasmid features using sequence analysis | |
Pooled Library Amplification | Amplify pooled-plasmid libraries | |
RNA Extraction Without a Kit | Extract RNA from tissues or cells without a commercial kit |

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 | |
pLKO.1 - TRC Cloning Vector | Cloning protocols for using the pLKO.1 vector, a backbone used by the RNAi consortium for targeting human and mouse genes. |

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 | Watch the Video! |
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 |