i'am working on Helicobacter pylori and i'm trying to get a pure colony but it's too difficult!
i wanna know the best sample and conditions for bacteriological isolation of H.pylori.
You can use selective media for bacteria Isolation, please see in attach file.
I used selective media but the isolation rate was very low
"wanna" is a vulgar version of "want to"
Try the enrichment step.
Growth in selective enrichment broths. H. pylori Special
Peptone broth (HPSPB), which contained the same ingredients as
HPSPA except the agar, was supplemented with various concen-
trations and combinations of antibiotics to create a selective en-
richment broth for isolating H. pylori. In each trial, six Erlenmeyer
flasks containing 96 ml of HPSPB without antibiotics and six
flasks containing HPSPB with various levels of antibiotics were
inoculated with H. pylori (ATCC strains 43504, 43629, or 43579).
For the first trial, the antibiotic combination tested was vanco-
mycin (10 mg/liter), amphotericin B (10 mg/liter), cefsulodin (10
mg/liter), and trimethoprim (10 mg/liter). Samples were collected
from the broths after 24 and 48 h of incubation, and 0.1 ml of
appropriate decimal dilutions was surface-plated on HPSPA, pro-
viding a total of 12 replications for each HPSPB antibiotic for-
mulation. After inoculation, all plates were incubated in a micro-
aerophilic atmosphere for 4 days at 378C, and the colonies were
counted. Colonies from each replicate were confirmed as H. pylori
as described above. The same methodology was used for three
subsequent trials, in which additional antibiotic combinations
were compared.
I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...
05 August 2024 6,787 4 View
Please address the best way to drop a plasmid. Background: I have a "bait" plasmid resistant to kanamycin and a "prey" plasmid resistant to carbenicillin. After many rounds of streaking on...
25 July 2024 2,532 3 View
I am cloning an overexpression plasmid with my protein of interest tagged with mScarlet. After transforming my ligated product into DH5α bacteria and plating on LB agar, I noticed colonies with a...
22 July 2024 5,953 6 View
I have done the same procedure with BL21 and pLysE but i got colonies but multiple times on rosetta i did but not getting. can anyone explain what would have gone wrong?
12 July 2024 9,904 1 View
How can we measure their colony diameter?
12 July 2024 7,027 0 View
Get too many colonies, which are mostly non-transformants when tested by colony PCR
11 July 2024 6,775 3 View
I am working on cloning of a gene fragment inside my vector everything we tried to do correctly like making insert and vector with utmost care but after ligation the morphology of colonies seems...
11 July 2024 9,010 3 View
The recharge of deep sandy groundwater is slow. Does the high level of nitrate promote bacteriological activity knowing that it requires a high level of O2? Thanks
10 July 2024 9,883 5 View
Hi. I have brightfield images of stem cells. I need to count the total number of cells/image. Some cells are individual and some cells are in colonies. Using average cross-sectional area of...
05 July 2024 4,197 1 View
Hello, recently i am an issue swith cloning where I am using the subclone method to insert my gene length is 1.5kb and my vector(pMAL-c6T) is 5kb I got the colonies which i screened through the...
04 July 2024 7,977 2 View