What to do when a plasmid comes for you: agar stab to plate to glycerol stock, miniprep & sequencing
I ordered a bunch of plasmids, and many of them come as agar stabs, where they've taken bacteria with the plasmid and stabbed it into a tube of agar. So it's like a Petri dish in a tube. My gameplan: streak a plate isolate a colony grow a 5mL overnight culture (LB + appropriate antibiotic) make a glycerol stock -- take 500μL of that culture & add to 500μL sterile glycerol; invert to mix & store in -80°C freezer - should be good for years if you avoid freeze-thaws miniprep the rest of the overnight culture to isolate the plasmid send the plasmid for sequencing if the sequence checks out, do whatever the heck you got it for! (subclone into a different vector, transform into expression cells, etc.) blog form: https://bit.ly/plasmid_prepping more on cloning cells: blog form: https://bit.ly/dh5alpha ; YouTube: • DH5α strains and a PSA to sequence your pl... more on expression cells: https://bit.ly/bl21_rare_codons ; YouTube: • Bacterial expression strains - esp BL21 st... more on molecular cloning: http://bit.ly/molecularcloningguide ; YouTube: • Molecular cloning overview - techniques & ... more on transformation: http://bit.ly/transformheatshock ; YouTube: • Bacterial transformation - chemically comp... more on T7 RNAP: http://bit.ly/t7rnap more on mini preps: http://bit.ly/minipreps ; YouTube: • Miniprep science - what goes on in each st... more on glycerol solution-making: • Making a 80% glycerol stock solution - a s... some tips for pipetting & working with viscous solutions: blog: https://bit.ly/pipetting_problems ; YouTube: • Pipetting viscous liquids - reverse pipett... random practical lab tips & tricks: https://bit.ly/lab_tricks_page link to the plasmid I was talking about: https://www.addgene.org/174866/ helpful guide from addgene: https://www.addgene.org/recipient-ins... more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 http://bit.ly/2OllAB0 or search blog: http://thebumblingbiochemist.com #scicomm #biochemistry #molecularbiology #biology #sciencelife #science #realtimechem

Random tips for working with DNA/RNA spin columns (like those in miniprep or PCR purification kits)

Extracting Plasmid DNA: How To Do a Miniprep

Molecular cloning overview - techniques & workflow
![Od Wielkiego Wybuchu, przez Dinozaury, po Człowieka - Historia Życia na Ziemi! [Podcast Historyczny]](https://i.ytimg.com/vi/ANgLZdszmVI/hqdefault.jpg?sqp=-oaymwEjCNACELwBSFryq4qpAxUIARUAAAAAGAElAADIQj0AgKJDeAE=&rs=AOn4CLBG2FoVhTbmicjlctCRo1plCwYKVw)
Od Wielkiego Wybuchu, przez Dinozaury, po Człowieka - Historia Życia na Ziemi! [Podcast Historyczny]

pGLO Bacterial Transformation Lab

What do tech pioneers think about the AI revolution? - The Engineers, BBC World Service

But what is quantum computing? (Grover's Algorithm)

How do I get started with Oxford Nanopore sequencing?

Making the stinkiest chemical known to man

How Microscopic Hunters Get Their Lunch

How to Harvest Plasmid DNA with Minipreps

AlphaFold - The Most Useful Thing AI Has Ever Done

BIOLOGY explained in 17 Minutes

Bacterial Transformation

But what is the Fourier Transform? A visual introduction.

Cloning cells (e.g. DH5α) vs. expression cells (e.g. BL21) & features of each

How to Make Chemically Competent E. coli

Plasmid design (bacterial expression vector)

Lecture 40 : Bacterial Culture for Plasmid DNA Isolation

