(r4) ProtocolsCulturingSnodgrassellaAlvi < Lab

---++ Working with _Snodgrassella alvi_

_Snodgrassella alvi_ is a member of the honey bee (_Apis mellifera_) gut core microbiota that can be cultivated _in vitro_. Its isolation and characterization were recently carried out by [[https://wkkwong.com][Waldan Kwong]] in the laboratory of [[http://web.biosci.utexas.edu/moran/][Dr. Nancy Moran]] and are described in depth in [[http://ijs.microbiologyresearch.org/content/journal/ijsem/10.1099/ijs.0.044875-0][here]].

---+++ _In vitro_ culture conditions

---++++ Plate Growth
 _Snodgrassella alvi_ strain wkB2 (type strain) is generally cultured at 35°C and 5% CO<sub>2</sub>. It grows robustly on agar plates after 48 hours on the following media: 
   * Heart Infusion Agar
   * Heart Infusion Agar supplement with Sheep's Blood
   * Columbia Agar supplement with Sheep's Blood (CBA)
   * R2 Agar

---++++ Liquid Media
 _Snodgrassella alvi_ can be cultivated in the following liquid media (same general culture conditions as above, without shaking): 
   * Insectagro (proprietary media)
   * Salt-free LB
   * R2 Liquid Media

---++++ Conjugation
 _Snodgrassella alvi_ can be conjugated using _E. coli_ donor strains and mobilizable rsf1010-based plasmids. Refer to the Bee Microbiome Toolkit paper for full description of working with this species. <Br>
Perform conjugation as described here: [[ProtocolsConjugation][Conjugation]], with the following specifics: use 10:1 ratio of _S. alvi_ to donor _E. coli_ pooled on CBA plates. Incubate overnight, then resuspend and plate on selective CBA plates.
Barrick Lab > ProtocolList > ProtocolsCulturingSnodgrassellaAlvi
Topic revision: r4 - 2018-10-25 - 19:51:07 - Main.SeanLeonard