SEA-PHAGES | Phage isolation in winter--any differences?

Link to this post \| posted 05 Feb, 2021 18:26
kmaclea	Dear Colleagues We are running phage discovery in the winter for the first time. Our first go-round we have a lot of success with direct isolations out of the box. This might have been an outlier leading us astray, but over the last couple of weeks we only had one successful run with direct isolation. We have started enriched isolation but still wondering if we can do better with our direct isolation or if winter in New Hampshire is just going to be more challenging. Anyone have thoughts? Kyle – Kyle MacLea Associate Professor, University of New Hampshire at Manchester kyle.maclea@unh.edu +1 603-641-4129

Link to this post | posted 05 Feb, 2021 18:26

Dear Colleagues

We are running phage discovery in the winter for the first time. Our first go-round we have a lot of success with direct isolations out of the box. This might have been an outlier leading us astray, but over the last couple of weeks we only had one successful run with direct isolation. We have started enriched isolation but still wondering if we can do better with our direct isolation or if winter in New Hampshire is just going to be more challenging.

Anyone have thoughts?

Kyle

–
Kyle MacLea
Associate Professor, University of New Hampshire at Manchester
kyle.maclea@unh.edu +1 603-641-4129

Link to this post \| posted 05 Feb, 2021 18:27
kmaclea	Added: We started with A. globiformis and had no luck, and tried again with M. foliorum and also had no luck. – Kyle MacLea Associate Professor, University of New Hampshire at Manchester kyle.maclea@unh.edu +1 603-641-4129

Link to this post \| posted 08 Feb, 2021 01:21
viknesh	kmaclea Dear Colleagues We are running phage discovery in the winter for the first time. Our first go-round we have a lot of success with direct isolations out of the box. This might have been an outlier leading us astray, but over the last couple of weeks we only had one successful run with direct isolation. We have started enriched isolation but still wondering if we can do better with our direct isolation or if winter in New Hampshire is just going to be more challenging. Anyone have thoughts? Kyle Hi Kyle, From what I can tell based on findings from across the SEA, the success rate for direct isolation is generally low and inconsistent (unless you keep going back to the same/similar spot). I'll let faculty from the northern reaches of the country share their advice, but I know several who gather soil early (before the ground freezes, and store them in sealed ziplocks at 4C) or use soil from indoor pots. I personally have had success with both methods when isolating phage for M. foliorum, but only by enriched isolation. Good luck!

Link to this post | posted 08 Feb, 2021 01:21

viknesh

kmaclea
Dear Colleagues

We are running phage discovery in the winter for the first time. Our first go-round we have a lot of success with direct isolations out of the box. This might have been an outlier leading us astray, but over the last couple of weeks we only had one successful run with direct isolation. We have started enriched isolation but still wondering if we can do better with our direct isolation or if winter in New Hampshire is just going to be more challenging.

Anyone have thoughts?

Kyle

Hi Kyle,

From what I can tell based on findings from across the SEA, the success rate for direct isolation is generally low and inconsistent (unless you keep going back to the same/similar spot). I'll let faculty from the northern reaches of the country share their advice, but I know several who gather soil early (before the ground freezes, and store them in sealed ziplocks at 4C) or use soil from indoor pots. I personally have had success with both methods when isolating phage for M. foliorum, but only by enriched isolation.

Good luck!

Link to this post \| posted 09 Feb, 2021 16:10
kmaclea	Thank you for that, Vic! I've got my fingers crossed for the enriched right now, we should know how our first run went soon. I had avoided getting soil before the frost since the guide had suggested fresh was best. I did get a good direct isolation from a place we isolated from last year so I do think your words about going back to the same spot could be prescient. Regardless–thank you! Hopefully some good success soon. Thanks! Kyle – Kyle MacLea Associate Professor, University of New Hampshire at Manchester kyle.maclea@unh.edu +1 603-641-4129

Link to this post | posted 09 Feb, 2021 16:10

kmaclea

Thank you for that, Vic! I've got my fingers crossed for the enriched right now, we should know how our first run went soon.

I had avoided getting soil before the frost since the guide had suggested fresh was best. I did get a good direct isolation from a place we isolated from last year so I do think your words about going back to the same spot could be prescient.

Regardless–thank you! Hopefully some good success soon. Thanks!

Kyle

–
Kyle MacLea
Associate Professor, University of New Hampshire at Manchester
kyle.maclea@unh.edu +1 603-641-4129

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Phage isolation in winter--any differences?