Robyn Raven

10 Questions 4 Answers 0 Followers

Questions related from Robyn Raven

How do I clone my protein in the 5'-3' orientation?

I have a question regarding cloning. I want to amplify a gene from a template plasmid, which I already have. The sequence reads -5’ TTA CDS CAT -3’ The reason the sequence is reversed is because...

01 June 2015 1,558 3 View

How do I bioinformatically identify secreted protein gram positive Bacteria?

I am looking into trying to characterise a protein secreted from a gram positive bacteria. I have started by aligning the sequence to a number of strains of the particular species to see which...

10 March 2015 2,406 5 View

What is the best way to prove interaction between two proteins?

I am trying to carry out experiments which will prove the interaction between two different proteins. One protein is present in the cytoplasm and is thought to be secreted into the environment,...

06 March 2015 627 7 View

Do you always have to include an RBS Site in your primer when cloning a gene?

I want to clone a gene into a vector and then western blot for the production of my protein after inducing expression. Do I need to include a RBS site sequence in my forward primer or will a...

03 March 2015 9,225 3 View

How do I bioinformatically identify secreted protein gram positive Bacteria?

I am looking into trying to characterise a protein secreted from a gram positive bacteria. I have started by aligning the sequence to a number of strains of the particular species to see which...

03 March 2015 5,188 6 View

What is the best way to prove interaction between two proteins?

I am trying to carry out experiments which will prove the interaction between two different proteins. One protein is present in the cytoplasm and is thought to be secreted into the environment,...

03 March 2015 4,001 8 View

How long do transformed Staphylococcus aureus cells take to grow?

I have electroporated staphylococcus cells with a plasmid using the COL and RN6390 strains and plated onto TSA Chloramphenicol and incubated at 37degrees, however after 4 days I have no growth....

01 March 2015 6,279 5 View

Inducing the proteins when carrying out bacterial two hybrid experiments?

I have cloned my genes into the p18 and p25 vectors and transformed them into BH101 cells. And plated them onto MacConkey (deficient in lactose) agar plates supplemented with 1% maltose, I am...

26 February 2015 7,230 1 View

How do I differentiate between my target protein and a protein present in Staph which both bind to anti-HA antibody?

I am working with staphylococcus aureus in the lab. I have cloned a target gene with a HA epitope tag and analysed the secretion of this target protein in the supernatant by western blotting using...

02 February 2015 4,886 6 View

Inducing the proteins when carrying out bacterial two hybrid experiments?

I have cloned my genes into the p18 and p25 vectors and transformed them into BH101 cells. And plated them onto MacConkey (deficient in lactose) agar plates supplemented with 1% maltose, I am...

02 February 2015 3,499 1 View