Chandrakala Gowda

8 Questions 19 Answers 0 Followers

Questions related from Chandrakala Gowda

Protein expressed in Deuterated M9 Media goes into pellet ?

Dear All, I tried to express a protein in a modified M9+ (Deuterated) media as suggested in “Cai, M., Huang, Y., Yang, R., Craigie, R., & Clore, G. M. (2016). A simple and robust protocol...

09 September 2018 9,564 8 View

Prodrg topology files, docking - can anyone help?

PRODRG server sometimes assigns CR61 for aromatic carbons and sometimes assigns CR1, CR0 for my ligand. What is the difference between the two? Where can I get more info on the atom types used in...

08 August 2014 5,085 5 View

Is it essential to have a M9 primary?

I follow the following protocol to grow my bl21de3/bl21de3-plysS 1. 10ml of MDAG-135 primary media to be inoculated with a single colony. (6 to 9 hours upto O.D = 0.6 to 1.0) 2. 100ml M9 to which...

01 January 1970 2,357 2 View

Very slow cell growth in M9 media with a generation time of roughly 4hours?

I have a situation with bacterial growth in M9 media with two plasmidsWild Type (WT) and Mutant. BL21DE3-PLysS cells. procedure followed is as follows 1. Incoulated 5ml LB primary with a single...

01 January 1970 6,502 6 View

BMRB submission of pdb structure obtained from cyana ?

My question is basically for someone who is familiar with CYANA software and have successfully submitted a pdb file calculated using NMR restraints using CYANA I am trying to execute the macro...

01 January 1970 2,846 0 View

Does anyone know the log-phase window for M9 media?

I ask this because if M9 media grows to a OD of 2 or 3 in the overnight culture are the cells in the log phase or stationary phase? my cells (bl21de3-plysS) are finding a hard time to grow in the...

01 January 1970 8,794 0 View

BL21DE3-PLysS cells give a slimy and sticky pellet...and have a pale wood yellow color?

BL21DE3-PLysS cells give a slimy and sticky pellet...and have a pale wood yellow color? Is it ok to have a slimy cell pellet? I suspect probably a few cells (may be around 10 % or so) lyse during...

01 January 1970 2,053 9 View

Cell clumps/floating debris found in primary M9 overnight culture?

I inoculate my 100ml M9 culture with cells from a 10ml MDAG - 135 media of 8 hours growth. After overnight incubation at 37degrees I see a good growth of the cells upto OD 2.0 but with also large...

01 January 1970 3,813 2 View