Are cloned plasmids less efficient in transfection?

Are pDNA that are cloned with the genes of interest less efficient? are there any drawbacks? Any drawbacks in relation with lipofectamine or using them with HeLa or HEK 293?

02 March 2016 1,795 6 View

Expressing GFP in HEK and HeLa. no ECL results?

I have been working with GFP that I have exprrssed in HEK and HeLa cell lines. On transfer I can only see the marker on the nitrocellulose. I cannot even see the positive control on the...

02 March 2016 574 5 View

Limitations and failures in western blotting and ECL?

If anyone could please suggest me research papers wherein western blotting and ECL have failed to produce any results. I have been trying to express GFP in HeLA and HEK and on transfer I can only...

02 March 2016 6,138 0 View

Disadvantages of green fluoroscent protein (GFP)

11 December 2015 7,845 1 View

Can anyone please tell me two old and current techniques used to diagnose scarlet fever or group A streptococcus in labs?

Scarlet method diagnostic methods

01 February 2015 9,754 6 View

Which microbes are good at forming biofilms and what morphological elements make them so?

Biofilm

01 February 2015 3,144 1 View

What is the exact mechanism of the antimicrobial effect of local anesthetics?

The antimicrobial effect of local anesthetics has been known for more than 50 years, but it still seems to have an unclear mechanism of action. which bacteria? which concentration? etc etc. ....

27 February 2021 4,450 2 View

Uncertianiny in using dry or wet basis in mixing a sediment with fuel oil?

Hello I'm currently working on a topic related to studying the anaerobic biodegradation of TPH in marine sediments. In this work, sediment should be mixed with fuel oil to a concentration of 20 g...

24 February 2021 9,839 3 View

Advice on growing lactobacillus species in the lab?

Hello everyone, For part of my PhD project, I'm hoping to grow some lactobacillus species in the lab so that I can create standard curves for a qPCR quantification of a faecal sample. However, I...

24 February 2021 3,157 6 View

I have a problem with kanamycin - why the kanamycin doesn't affect?

Hi everyone, I need to expression of GFP with pET28 plasmid in E. coli BL21(DE3) plysE. Transformed cells have kanamycin and chloramphenicol resistance. Normally, only transformed cells should...

22 February 2021 4,032 4 View

What are the best cations for preparing competent cells, Ca2+ or Mg2+?

I have been reading various articles and most state that treatment with CaCl2 produces higher efficiency of transformation than MgCl2. Can anyone explain why Ca2+ is better than Mg2+ at...

16 February 2021 8,519 5 View

Can 5-FOA Diffuse Through Agar?

I am preparing to do a gene knockout in the filamentous fungi Coccidioides posadasii targeting pyrG/URA3 so that 5-FOA can be used for selection pressure instead of traditional antibiotic...

16 February 2021 169 4 View

Would using an asRNA for repressing transcription of a high expression Listeria monocytogenes non-coding RNA harbored in a plasmid be a viable option?

Hello all, I am interested in deleting a highly expressed non-coding RNA located on a Listeria monocytogenes plasmid. Currently, there are not any optimized gene deletion methods for Listeria...

09 February 2021 4,994 1 View

Review Papers regarding transcriptional regulations & promoter types/classes in Gram positive bacteria?

Dear fellow researchers, Could you please suggest me excellent review papers discussing transcriptional regulations and/or promoter classes in Gram positive bacteria? Thank you so much for your help!

08 February 2021 6,909 3 View

Flexibility of RNAseq?

When you use RNAseq I understand that it must follow certain conditions when you use it. One of them is the insertion of a polyA tail. The insertion of the polyA must include then...

05 February 2021 3,770 4 View

How do microbes tell the presence of an inorganic minaral in the soil?

Due to their ubiquitous nature, there is a tendency of the presence of microbes in a given mineral(s). For instance, if microorganisms that love the of pH less than 6 in the pH value, is isolated...

03 February 2021 4,630 10 View