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2X smeg top agar
| Link to this post | posted 19 Sep, 2022 18:56 | |
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Hi! Is there is a reason the smeg top agar is prepared at 2X, and then diluted to 1X? I'm wondering if I can prepare it as a 1L volume, autoclave, and then aliquot into smaller bottles (as with the PYCa top agar). We are phage-hunting with smeg for the first time this semester. Thanks! Liz |
| Link to this post | posted 21 Sep, 2022 17:09 | |
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edoddmoh@uottawa.ca Hi Liz, Part of the reason top agar is prepared at 2x is so that experiemnts that require the use or large samples of phage (in buffer) do not overly dilute the nutrient in the medium. For our general PHAGES protocols, you should be fine preparing top agar at 1x. 7H9 medium has lots of salts in it, and calcium chloride tends to precipitate out over time. For this reason, calcium chloride is only added before use. Even if you prepare 1x top agar, you might want to only add calcium chloride before use. If you do add it early, please let us know how long it takes for the precipitates to form so that we can share with others. Vic |