Barrick Lab :: ProtocolsWorkingWithSerratiaMarcescens

---++ Working with _Serratia marcescens_

_Serratia marcescens_ is a gram-negative pathogenic bacteria known for its distinctive red pigmentation. While it is not found in the microbiome of healthy honey bees (_Apis mellifera_), it is capable of living in the gut of sick or aging bees. _S. marcescens_ can be cultured _in vitro_.

---+++ _In vitro_ culture conditions

Both strains of _S. marcescens_ used in the Barrick lab, N10A48 an KZ2, grow robustly in LB after 24 hours at 30°C. _S. marcescens_ will form colonies on LB agar after 24 hours at 30°C. _S. marcescens_ has some natural tolerance for spectinomycin. When working with spectinomycin-resistant _S. marcescens_, the working concentration of spectinomycin should be increased from 60 mg/mL to 180 mg/mL for this reason.

---++++ Isolating plasmid DNA ("Miniprep")

 _S. marcescens_ N10A48 and KZ2 do not pellet well after lysis when isolating plasmid DNA. Instead of forming a neat pellet, cell debris forms a gelatinous blob that makes it difficult to remove the supernatant. Increasing the speed and time of the centrifugation step to 15000x for 20 minutes helps form a more compact pellet. 

-- Main.StrattonGeorgoulis - 14 Dec 2017

Barrick Lab > ProtocolList > ProtocolsWorkingWithSerratiaMarcescens

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Topic revision: r1 - 2017-12-14 - 22:02:46 - Main.StrattonGeorgoulis