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ProteoCool n°5
(E.coli high cell density media overview - 18/11/2018)
(E.coli high cell density media overview - 18/11/2018)
ProteoCool n°7
(High cell density E.coli protein expression using EnpressoB media - 08/12/2018)
Detailed Proteocools in pdf are available at:
https://www.researchgate.net/publication/329505526_Ecoli_high_cell_density_protein_expression_in_24-Deep_Well_using_Enpresso_B_media
https://www.researchgate.net/publication/329505518_E_coli_high_cell_density_protein_expression_in_shake_flask_using_Enpresso_B_media
https://www.researchgate.net/publication/315999127_Multiwell_based_protein_expression_and_solubility_test
ProteoCool n°22
(Overview on Tools for recombinant protein expression in E.coli - 17/07/2019)
ProteoCool n°29
(Expi 293: A Simple and powerful mammalian expression system - 28/04/2020)
ProteoCool n°30
(Uniform 15N, 13C and 2H protein labeling using E.coli and more - 23/05/2020)
(Uniform 15N, 13C and 2H protein labeling using E.coli and more - 23/05/2020)
ProteoCool n°32
(High cell density E.coli protein expression using YGR media -12/08/2020)
(High cell density E.coli protein expression using YGR media -12/08/2020)
Please
in case you found some of this ProteoCool able to improve your work: Leave a comment.
Your feedback could be useful to the others
Thanks.
Manuele
Hi there! Thanks for your videos, they are very interesting. I am interested in the YGR media recipe and protocol. Is there any reference for this particular media, describing the recipe that you show in the video? Do you have the detalide protocol to use it? Thanks again for your time.
ReplyDeleteHello! Ciao :-)
DeleteThanks a lot for your nice comments!
To my knowledge the only paper citing the YGR (named HTMC there)is:
Scietti et.al Exploring host-pathogen interactions through genome wide protein microarray analysis. Sci Rep. 2016 Jun 15;6:27996.
At the moment I do not have ready any video ready for YGR media but never say never :-)
However the protocol for auto-induction is quite simple:
Dilute a LB pre-culture 1:100 in YGR and let it to growth for about 40h at 30°C -160 rpm. For this media, as for the Enpresso, the culture volume need to be optimized on the basis of the shake flask. For example for a 250 ml buffled shake flask with cup vented, 50ml of culture is the optimal volume.
good luck
Manuele
Gracie mille!
DeleteI can see that they don't add glucose though... As for your video, I understand that glucose is only an early repressor of the T7 promoter, wich will allow an initial bacterial growth without induction and, when it is consumed, induction will take place. So, the addition of glucose will just create an "initial growth window" similar to what it is usually done in standard procedures before adding the IPTG. Is that right?
Prego! You are welcoome!
DeleteSincerelly i used for several years the pubblished version an it works quite well. AT one point one my colleague, that was one of the scientist that settesd-up the HTMC suggested to me to add this small amount of glucose, expecially for shake flask production, that can be important expecially for toxic proteins and i'm currently using this version, but in my experience also the pubblished version work quite well.
ciao
Manuele
This was incredibly an exquisite implementation of your ideas ausjuice
ReplyDeleteThank you for sharing your knowledge. This is so awesome and helpful for young scientists.
ReplyDeleteI Like to add one more important thing here, The protein expression market was valued at USD 1,654.0 Million in 2017 and is expected to grow at a CAGR of 11.5% to reach US$ 3900 Million in 2025.
ReplyDelete