Low melting point agarose

[@thebumblingbiochemist]

You don’t want the boots on *all* the “feet” because you still need plenty of shoeless feet available to H-bond or else you can’t form a gel in the first place. The modifications are added using organic chemistry (yay o-chem!, I miss you! ). Usually, a base is used to steal an -OH’s “H” so the O becomes a strong nucleophile (positivity-seeker) & attacks the “boot” you want to latch on. More on this sort of thing here: bit.ly/nucleophilefiles    ⚠ LMPA also takes a lot longer to set than normal agarose. I learned this the hard way in undergrad - I was accidentally using LMPA instead of NMPA because I didn’t realize the bottles were different (they both said agarose, right?). My gels were taking forever to set & were super fragile. it was my 1st time(s) running them, so I thought that was normal. It was such a relief when I found out it wasn’t! Morals of the story - when using LMPA, expect to wait longer & always read bottles closely!  After you microwave it, let it cool to ~60°C before pouring. Once it cools to room temp you can let it finish setting in the cold room. Also, some people say to run the gel with cooled buffer so it doesn’t overheat and melt the gel (along these lines, don’t go cranking up that voltage). Note that fragments will run differently than in normal melting point agarose. They’ll run faster so you should run at a lower voltage More info in these product manuals: www.goldbio.com/documents/3976/Low%20Melt%20Agarose%20Gel%20Preparation%20Protocol.pdf www.avansbio.com/Upload/ProductDownload/00000009/LMP%20Agarose%20Gel%20Preparation%20Protocol.pdf and this CSH Protocol: Recovery of DNA from Low-Melting-Temperature Agarose Gels: Organic Extraction by Michael R. Green and Joseph Sambrook cshprotocols.cshlp.org/content/2020/3/pdb.prot100461.full a video of mine on more things about agarose: kzbin.info/www/bejne/l4fNaZuehNqjgtU   more about agarose gel electrophoresis: bit.ly/agarosegelcompare more about crush & soak gel extraction: blog: bit.ly/gel_extraction_crush ; KZbin: kzbin.info/www/bejne/mJKxoZeua9mrhdE        more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 bit.ly/2OllAB0 or search blog: thebumblingbiochemist.com

[@jackmackerel4151]

What about low EEO agarose? Is low melting point always better for gel extraction? My lab has standard, low EEO and low melting point agarose on the shelf. Is low EEO or low melting point preferred when doing gel extraction for downstream processes?

[@thebumblingbiochemist]

I hadn't heard of EEO agarose , sorry