ESO wikiadmin: Created page with "{{Protokoll-Mix|Inhalt=Gonococcal strain which is Opa-postive /Pili positive PPM medium (15g proteose pepton, 5g NaCl, 4g KH 2PO4 1g K2HPO4, 1g soluble starch, ad 1l H20 ph: 7.5) + vitamin mix cloned and sequenced analysed gonococcal DNA which contains the spectinomycin resistance cassette and the Neisserial DNA uptake sequence (DUS) (plasmid DNA}} == Procedure == * Ngo strain (O+/P+) were grown overnight (37°C) on GC agar plates in a humid atmosphere with 5% CO 2 * N..."

2025-06-02T07:49:19Z

Created page with "{{Protokoll-Mix|Inhalt=Gonococcal strain which is Opa-postive /Pili positive PPM medium (15g proteose pepton, 5g NaCl, 4g KH 2PO4 1g K2HPO4, 1g soluble starch, ad 1l H20 ph: 7.5) + vitamin mix cloned and sequenced analysed gonococcal DNA which contains the spectinomycin resistance cassette and the Neisserial DNA uptake sequence (DUS) (plasmid DNA}} == Procedure == * Ngo strain (O+/P+) were grown overnight (37°C) on GC agar plates in a humid atmosphere with 5% CO 2 * N..."

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{{Protokoll-Mix|Inhalt=Gonococcal strain which is Opa-postive /Pili positive
PPM medium (15g proteose pepton, 5g NaCl, 4g KH 2PO4 1g K2HPO4, 1g soluble
starch, ad 1l H20 ph: 7.5) + vitamin mix
cloned and sequenced analysed gonococcal DNA which contains the spectinomycin resistance cassette and the Neisserial DNA uptake sequence (DUS) (plasmid DNA}}

== Procedure ==

* Ngo strain (O+/P+) were grown overnight (37°C) on GC agar plates in a humid atmosphere with 5% CO 2
* Next day colonies were resuspended in 1ml PPM medium/Vitamin mix for 2 hours at 37°C with an agitation at 220 rpm
* The fresh suspension was measured at OD550 nm and afterwards diluted to OD550 nm of 0.1
* 1 ml of this gonococcal suspension is then incubated with 1 μg of the gonococcal plasmid DNA and incubated for 6 hours at 37°C with an agitation of 100 rpm
* The transformed bacteria are than seeded (undiluted, 1:10 and 1:100 diluted) on selective GC- agar plates containing spectinomycin and incubate for 2 to 3 days

Transformation of Neisseria gonorrhoeae - Revision history