Sample preparation: Metabolite extraction (tutorial 3/5)

Рет қаралды 23,169

European Molecular Biology Laboratory (EMBL)

4 жыл бұрын

The metabolome of a biological system can change within a few seconds or minutes. Sample preparation is one of the most critical parts of a successful metabolomics experiment.
Sample collection, processing and storage can affect concentrations of metabolites changes, resulting in an inaccurate data.
Here we demonstrate a step-by-step procedure for aqueous methanol extraction of polar metabolites from HeLa cell culture. This general protocol can be adapted for other samples considering: 1) type of biological matrix 2) sample collection 3) quenching of cellular metabolism 4) extraction and recovery of metabolites 5) drying 6) storage and shipment.
Read more about protocols via www.embl.de/mcf/metabolomics-....

Пікірлер: 23

@lizhang2849 3 жыл бұрын

Today I have 90 samples prepared for targeted metabolomic assay. thank you.

@shakirshamas6286 Жыл бұрын

superb, really awsome and informative lecture

@Etatdespritdecroissance 6 ай бұрын

GREAT video. Thanks.

@faithogbonna5484 2 жыл бұрын

Please what did you use to aspirate? Or can you please explain the aspiration part?

@violettracey 10 ай бұрын

Thanks!

@emmanuelrios6832 3 ай бұрын

super. Thank you

@faithogbonna5484 2 жыл бұрын

Please can you explain the quenching procedure? Aspiration was done . What was used for aspiration? Am currently working on extraction of metabolites from Enterococcus species and I need more guides

@shakirshamas6286 Жыл бұрын

same question and same procedure and guidance I needed. i am working on primary metabolites of chickpeas

@leewilliam3417 4 ай бұрын

Thankyou 😊

@azizabed7692 3 жыл бұрын

Hi, Thanks for this video. Please I have 2 questions: 1- I why you stored the extraction solution for 1 hour before use? 2- What is the red solution? Is it bacteria cells with its extracts? Could you please please tell me more about the bacteria cells sources , preparation , how you stimulate them to excrete secondary metabolite? Thanks

@EMBLorg 3 жыл бұрын

Information about the protocols can be found here: www.embl.de/mcf/metabolomics-core-facility/protocols/index.html including contact details for the EMBL Metabolomics Core Facility, if you have further questions.

@PrasadPhapale 3 жыл бұрын

Hi Aziz, 1) the storage is required to have cold methanol for extraction. The cold methanol is necessary for quenching the metabolism before extraction 2) This protocol is for adherent mammalian cells; the red color is from DMEM medium in which cell are grown.

@Powerofou 2 ай бұрын

I put my cell lysate in -80. Can I use them for metabolomic? Too late to add methanol to quench metabolism?

@purplerain365 4 жыл бұрын

hi, can I get a softcopy of this protocol of metabolite extraction for my final year project? Thank you.

@EMBLorg 4 жыл бұрын

Please contact the Metabolomics Core Facility team, clearly stating the tutorial you are referring to and what you would like them to send you. You can contact them through the EMBL website: www.embl.de/mcf/metabolomics-core-facility/contact/index.html

@PrasadPhapale 3 жыл бұрын

Hi Jennifer, soft copy is available on our webpage under protocols. Thanks!

@jarrysc 3 жыл бұрын

Hi, Jenn, Did you find the protocols?, can we speak in a private conversation by mail... javiersanchezing@gmail.com, will write whoever worked with extraction of metabolites... I'm really trying to get the way to extract from Trichoderma sp. and I have actually lot of questions about.