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Viknesh Sivanathan posted in did you know you can do restriction digests in the microwave?
nic.vega posted in did you know you can do restriction digests in the microwave?
Patch Assay Help
| Link to this post | posted 15 Feb, 2022 16:15 | |
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Good morning. I have question concerning the patch assay. Each time I have performed the patch assay, there is growth on where the potential lysogen should be. Instead of having clearing on where my potential lysogen should be, I keep having growth. I am not sure what to do next. I have been following the protocol stated on 11.2 of the phage discovery guide. |
1.94Mb| Link to this post | posted 15 Feb, 2022 16:27 | |
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Hi! You should have bacterial growth! But if it is a lysogen, that bacterial growth should be releasing phage, so around the bacteria streak you will see a zone of lysis. You can see a tiny zone of lysis around A and D. But it is not easy to tell. What phage are you working with? debbie |
| Link to this post | posted 15 Feb, 2022 17:58 | |
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The bacteria that I am using in the research is Arthobacter golbiformis, and the lysate is called Basilisk. |
| Link to this post | posted 15 Feb, 2022 18:18 | |
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Next step, is to test the grow a culture from a streak on WT A. globi. (I would pick D.) From that culture, you will want to make a lawn and then test the phage for infection (by plating a serial 10-fold dilution) AND spinning some of the cells down and testing (by plating a serial dilution) the supernatant on the WT A. globi. Make senee? debbie |
| Link to this post | posted 15 Feb, 2022 18:32 | |
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I would just do the protocol of 11.3, right? |
| Link to this post | posted 15 Feb, 2022 20:11 | |
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yep! |