Link to this post | posted 06 Nov, 2025 21:50
NWCiowaSEAPHAGE	Here's an example of a phage stained with UranyLess in our lab. - sara

Link to this post | posted 06 Nov, 2025 21:50
NWCiowaSEAPHAGE	Here's an example of a phage stained with UranyLess in our lab. - sara

Link to this post | posted 06 Nov, 2025 21:50
NWCiowaSEAPHAGE	Here's an example of a phage stained with UranyLess in our lab. - sara

Link to this post | posted 06 Nov, 2025 16:17

NWCiowaSEAPHAGE

We have recently resumed imaging our phages with TEM. We are a small insitution without a radioactive license so cannot use uranyl acetate. We used UranyLess stain and Lead Citrate enhancer (see photo attached. We put drops of each solution on Parafilm then floated grids on: HTL 10-20 minutes, water 1 min, Uranyless 1 min, water 1 min, Lead Citrate 2 min, water 1 min. I've also attached an image to a reply to this post that we got using these reagents and this protocol (full transparency–it was one of our best images). - Sara Tolsma 2.35Mb

Link to this post | posted 11 Sep, 2025 18:18

NWCiowaSEAPHAGE

NWCiowaSEAPHAGE The attached spreadsheet reports on the host range of 12 phages: JacoRen57, DrLupo, Quesadilla, and Knocker (isolated on M. smegmatis), Challenger, RedRaider, BlueWizard, and KoBear (isolated on G. terrae), DoDo, IndiRoo, Exploradora, A3Wally (isolated on M. foliorum. Exploradora was able to infect M. smegmatis as well as M. foliorum, its isolation host. I've attached a data card to a subsequent post that includes experiments to confirm this. 643Kb

Link to this post | posted 11 Sep, 2025 18:14

NWCiowaSEAPHAGE

The attached spreadsheet reports on the host range of 12 phages: JacoRen57, DrLupo, Quesadilla, and Knocker (isolated on M. smegmatis), Challenger, RedRaider, BlueWizard, and KoBear (isolated on G. terrae), DoDo, IndiRoo, Exploradora, A3Wally (isolated on M. foliorum. Exploradora was able to infect M. smegmatis as well as M. foliorum, its isolation host. I've attached a data card to a subsequent post that includes experiments to confirm this. 1.89Mb

Link to this post | posted 06 Oct, 2024 00:55
NWCiowaSEAPHAGE	We're looking for protocols and reagents folks have used successfully for TEM that are not radioactive. Thanks in advance for any help you can give us!

Link to this post | posted 03 Jun, 2023 15:30

NWCiowaSEAPHAGE

We recently annotated two EM phages. The auto-annotations for both produced many many false positive genes. We saw reverse genes without BLAST hits without much coding potential completely overlapping forward genes with good CP and BLAST hits. We had to delete nearly half of the auto-annotated genes. Debbie added: Note when you review this in DNA maser, you will find the GeneMark has called all of the forward genes and Glimmer has called all of the reverse genes. this is based on how these 2 programs call this differently OR is it because there is a big open reading frame in the same location and the programs discriminate which one to pick differently. If you simply discard one of the sets of data, are you discarding possible genes. if all of these genes are in PECAAN, always good to see if you are throwing away anything with a functional…. Edited 06 Jun, 2023 02:05 521Kb

Link to this post | posted 14 Mar, 2022 15:10
NWCiowaSEAPHAGE	Thanks! Very helpful!

Link to this post | posted 04 Mar, 2022 16:25
NWCiowaSEAPHAGE	Mitch, Sorry! The numbering has changed since we annotated. Here is its sequence: MPVTPGDVTGARIRWRERSAVRHYDGDPEPSYREDVDEEDGPFKLGAEIGRGTDDDDTPIFSVRMKGSFTKPDAQVRIDVEVVFKIDSGEEVDQAFIQKHAMPYVFGYVRGGFTDACRSVGLPGWMIPMVDLAADIQYEDA - sara