[DIYbio] Phage assisted DNA distribution

Hey guys!

I've been working on a few things lately, one of which is an easy transformation protocol, mainly for distributing DNA easily. One of the problems I saw was that every time I recieved DNA, I had to make comp cells, which as you probably know takes some time and I was pretty lazy, mostly since my comp cells are only really good once, albeit a little less efficient. Not only that, but most people send DNA in centrifuge tubes or on paper, and for the fliter paper method I've had some problems in the past recovering plasmid. In addition, if I ever wanted to ship someone a plasmid I'd have to miniprep to make sure I give them "fresh DNA"

Off of the topic of distributing DNA, I was also thinking about starter kits. I've noticed the #1 thing beginners  mess up on is making competent cells. Whenever I've taught classes, here is were people mess up. When doing it at home, it's easy to get frustrated and quit because the first transformation with a start plasmid failed (like pGlo)


So, I came up with a possible alternative. Packaging a plasmid that expresses GFP into M13 phage particles. When I got M13KO7 from a neighboring lab (it was for a different project) they told me to just mix it with bacteria that have the F plasmid. (SS320 in my case). I simply mixed the phage and bacteria that were growing, and recovered (its a kan plasmid) and plated. Got tons of colonies. All you need for the M13KO7 to package your selected DNA instead of itself is a plasmid with the F1 origin, which is super common. (Even in pGlo!)

If this was adapted for ampicillin plasmids, all you'd have to do is mix with the phage with bacteria for a few minutes and plate. In fact, if you just put the phage on a filter disk and added that into some bacteria it'd work (there was a story of a guy who had really needed a secret phage from a lab, and they wouldn't give it to him, so he sent them a letter pleading for it, and they sent a letter back saying no. He dissolved the letter and recovered intact phage from the paper!). So, in general, recovery would be super easy, even for any DIYbiologist.

Next is distributing the actual DNA. Since E coli are constantly giving off phage (if infected with the helper plasmid, M13KO7) all you have to do is centrifuge the E coli and put the supernatant on filter disks. The entire registry of standard biological parts *could* be distributed like this for a fraction of the current cost (their standard plasmid doesn't have the F1 origin :( ). The entire library of parts could then be restored and saved since the amount of competent cells is no longer a problem. I am currently going to test this, I have pGLO and M13KO7 inside of Mach1s and some SS320 cultures going for testing this week.

However, I'd like some discussion on this, mainly with the disadvantages-

Advantages
Cheap
Easy to recover
Easy to distribute

Disadvantages
Possibly mutates quicker? (single stranded intermediate?)
Size limitation*
Lab receiving has to have F plasmid E coli
Contaimination issues***
(There would probably be a ton of contaimination since the phage are so infectious )

Anyone with experience have any comments on this? Or anyone in general, I'd like to know what you think about this system/idea of distributing parts to DIYbiologists
-Koeng