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How to make a tetracycline solution


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#1 slapolla

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Posted 25 March 2005 - 09:40 AM

I am making at 5 mg/mL soln of Tetracycline, and am using EtOH per our most recent Molecular Cloning manual...but I am having trouble getting all of the solid dissolved. Should I be using 70 % ETOH? Any thoughts/suggestions would be appreciated!!

#2 matiascs

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Posted 25 March 2005 - 12:56 PM

Ive made some Tet sol. a couple of times....be sure to use an analytical balance, and dissolve it in absolute ethanol (100%)....and shake it up like theres no tomorrow...itll take a while but it will go into solution...
Do not autoclave...its ready for use....
Cheers





I am making at 5 mg/mL soln of Tetracycline, and am using EtOH per our most recent Molecular Cloning manual...but I am having trouble getting all of the solid dissolved. Should I be using 70 % ETOH? Any thoughts/suggestions would be appreciated!!

<{POST_SNAPBACK}>


Mathias Casás Selves

Molecular Pathology Unit
Dept. of Biochemsitry
School of Medicine
Montevideo, Uruguay

#3 chandima

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Posted 29 March 2005 - 03:04 AM

i agree with matiascs. I do the same
chandima

#4 Julio-Claudian

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Posted 29 January 2009 - 05:54 AM

People,

Err... How do we know when to use water or ethanol? We have so much stocks here that I have yet to prepare any antibiotic solutions.

#5 mdfenko

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Posted 29 January 2009 - 07:53 AM

People,

Err... How do we know when to use water or ethanol? We have so much stocks here that I have yet to prepare any antibiotic solutions.


you use that in which it is soluble and stable and will not interfere with your purpose.
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#6 Julio-Claudian

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Posted 29 January 2009 - 04:25 PM

People,

Err... How do we know when to use water or ethanol? We have so much stocks here that I have yet to prepare any antibiotic solutions.


you use that in which it is soluble and stable and will not interfere with your purpose.


Alright, got it!

#7 molgen

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Posted 12 February 2009 - 09:24 AM

According to Lab-facts (Roche) make the stock in Et-OH and the dilutions in liquid culture.

#8 phage434

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Posted 13 February 2009 - 08:42 AM

I believe the confusion is that two forms of tetracycline are marketed, the compound, and its hydrochloride. Tetracycline is soluble in EtOH, but not water. Tet-HCl is soluble in water. Dissolving Tet-HCl in 50% EtOH is what we like, since it keeps the solution at -20C liquid.

#9 hanming86

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Posted 15 February 2009 - 08:34 AM

I believe the confusion is that two forms of tetracycline are marketed, the compound, and its hydrochloride. Tetracycline is soluble in EtOH, but not water. Tet-HCl is soluble in water. Dissolving Tet-HCl in 50% EtOH is what we like, since it keeps the solution at -20C liquid.


And from my experience Tet-HCL is not as soluble in Ethanol compared to water. Tet-HCL in Absolute EtOH is quite crappy in term of solubility.

Edited by hanming86, 15 February 2009 - 08:34 AM.

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#10 DNA

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Posted 12 March 2009 - 07:22 PM

Hi Guys,

I f we make tetracycline stock solution i absolute or 100% ethanol instead of water, dont u think that ethanol is dangerous/killer for E.Coli cells?

Yes i agree that not all tetracyline get dissolved , but i am still using it anyways by giving it a bit vortex.

Please advise.

Thanx.

#11 phage434

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Posted 12 March 2009 - 08:34 PM

The 1000x dilution you do when adding the stock to the medium makes the EtOH in the medium inconsequential.

#12 DNA

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Posted 12 March 2009 - 08:57 PM

The 1000x dilution you do when adding the stock to the medium makes the EtOH in the medium inconsequential.



Thank you very much phage434. I did not know about it that we can do it.

We have got Tet-Hcl and i would like to prepare it your way i.e 50% ethanol so it will remain in liquid form and no freeze thaw.

Cheers

#13 Resubmit

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Posted 25 August 2013 - 10:19 AM

In our lab, we dissolve Tetracycline-HCl in methanol to 10 mg/mL. The solution does not freeze at -20 C, and works well as a 1000x stock for use with E. coli.
(70% ethanol does also dissolve Tet-HCl well, however I've experienced precipitation after freezing the solution).



#14 El Crazy Xabi

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Posted 25 August 2013 - 05:03 PM

Tet in ethanol, Tet-HCl in MQ as basic rule

 

I made my stocks of Tet-HCl at 25 mg/mL without problem



#15 labtastic

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Posted 05 September 2013 - 04:29 AM

In my experience, tet will dissolve nicely in EtOH (or EtOH/H2O mixtures) but will eventually precipitate, especially at -20C. We dissolve our tet in DMF at a 1000x stock (25mg/ml). When my former boss told me to use DMF, I thought for sure it would have negative effects on the growth of my e.coli cultures, to the point where I bet him a 6-pack of beer and then did a growth curve of e.coli +/- pure DMF diluted 1:1000 in the media. Turns out I lost the bet- E. coli had no problems with the DMF. Plus the tet is very stable in DMF and doesn't freeze at -20C.






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