Was going to write "it's not that bad or hard", but it would be complete lie. It is hard and difficult and very expensive. But if one day you'll go in, you'll be amazed... You can make, for example, orchid, which can glow in dark only after 30C ambient temp. Or something else, limited by your fantasy only. CRISPR hardware and reagents also can be ordered and they are not even expensive (last time I had a look, guy was preparing for mass production of nano-syringe for DNA injection directly into nucleus). You can find polymerases and other stuff on ebay. Look for book "Molecular biology of cell, 7th edition" (older ones are also good, but well).
But again, it's insanely complex and time-consuming process (notice, I didn't include frustration and nerves :-) )
Funny part comes in, when every detail costs 10000x more than one step lower (Pipettes. PET pipette 100x costs 1$. Normal pipette 15 cents each. Look for dual channel pipettes which you'll need.. $800 starting price) (same goes for almost everything..) Hardware for sequencing is "not that expensive" (if compared to other things), but other things are just ...
np :) For preparations, I'd recommend going for YT video XVdw2IP1Smw . If you can somehow stand accent, he's very good at explaining how and why and when to add. P.S. Are you after only tissue culture or diving even deeper? (DNA modifications/sequencing/CRISPR/a bit of biochem/bioinfo?)
I mean, if your medium already contains EDTA as free acid (usually orchid mediums do this) and you'll be adding MnEDTA as Mn source (not sulfate, as it usually is added), just subtract mass of EDTA anion in MnEDTA from EDTAacid mass of medium.
Absolutely np. if you'll struggle finding some clean metal salts, let me know. I think I can help with them.
NaOCl you can make from Ca(OCl)2 and some sodium carbonate or sulfate or such.
In case you're worrying precipitating transition metal salts after time (Fe, Mn and Mo tend to do it, along with others). Try to have them already chelated with organics (EDTA/EDDHA/etc) (You can find them on phygenera) or just do them before making solution (quite annoying) (Just make sure chelating agent is not toxic and/or subtract anion mass from main mass, if you do them separately(in MS medium you got EDTA, so if you add MnEDTA, just add less amount of EDTA acid itself)
You also can try to use some UV-A lamp for desinfection, quite clean and convinient way, if it's not destroying reagents :)
And one last thing. In case you're struggling with pH, try to add buffers (MES is most popular, imho, but you can experiment/try other ones too).
P.S. Linux/unix system deployment engineer here :)
You're welcome. about other ingedients (which are not on phygenera) and/or if you need some professional hardware/reagents/books, try to look me up, Will be only glad to help even with sending them from EU to you :)
Just read your article. Wonderful, beautiful. Huge respects for building LFH by hand (still struggling with it, even after year of preparations.).
About chemicals you would need, go at https://phygenera.de/ . They got all vitamins, Agar, PGM's, etc (Also parafilm, you will love to use it for covering things.)
OK, last post and I stop spamming.
Was going to write "it's not that bad or hard", but it would be complete lie. It is hard and difficult and very expensive. But if one day you'll go in, you'll be amazed... You can make, for example, orchid, which can glow in dark only after 30C ambient temp. Or something else, limited by your fantasy only. CRISPR hardware and reagents also can be ordered and they are not even expensive (last time I had a look, guy was preparing for mass production of nano-syringe for DNA injection directly into nucleus). You can find polymerases and other stuff on ebay. Look for book "Molecular biology of cell, 7th edition" (older ones are also good, but well).
But again, it's insanely complex and time-consuming process (notice, I didn't include frustration and nerves :-) )
Funny part comes in, when every detail costs 10000x more than one step lower (Pipettes. PET pipette 100x costs 1$. Normal pipette 15 cents each. Look for dual channel pipettes which you'll need.. $800 starting price) (same goes for almost everything..) Hardware for sequencing is "not that expensive" (if compared to other things), but other things are just ...
All the best wishes and sorry for spam.
np :) For preparations, I'd recommend going for YT video XVdw2IP1Smw . If you can somehow stand accent, he's very good at explaining how and why and when to add.
P.S. Are you after only tissue culture or diving even deeper? (DNA modifications/sequencing/CRISPR/a bit of biochem/bioinfo?)
mmmmm.. I went after this medium: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/286/522/p6793dat.pdf
Works as charm, as people say.
I mean, if your medium already contains EDTA as free acid (usually orchid mediums do this) and you'll be adding MnEDTA as Mn source (not sulfate, as it usually is added), just subtract mass of EDTA anion in MnEDTA from EDTAacid mass of medium.
Absolutely np. if you'll struggle finding some clean metal salts, let me know. I think I can help with them.
NaOCl you can make from Ca(OCl)2 and some sodium carbonate or sulfate or such.
In case you're worrying precipitating transition metal salts after time (Fe, Mn and Mo tend to do it, along with others). Try to have them already chelated with organics (EDTA/EDDHA/etc) (You can find them on phygenera) or just do them before making solution (quite annoying) (Just make sure chelating agent is not toxic and/or subtract anion mass from main mass, if you do them separately(in MS medium you got EDTA, so if you add MnEDTA, just add less amount of EDTA acid itself)
You also can try to use some UV-A lamp for desinfection, quite clean and convinient way, if it's not destroying reagents :)
And one last thing. In case you're struggling with pH, try to add buffers (MES is most popular, imho, but you can experiment/try other ones too).
P.S. Linux/unix system deployment engineer here :)
You're welcome. about other ingedients (which are not on phygenera) and/or if you need some professional hardware/reagents/books, try to look me up, Will be only glad to help even with sending them from EU to you :)
Hi,
Just read your article. Wonderful, beautiful. Huge respects for building LFH by hand (still struggling with it, even after year of preparations.).
About chemicals you would need, go at https://phygenera.de/ . They got all vitamins, Agar, PGM's, etc (Also parafilm, you will love to use it for covering things.)
All the best..