Alia Mike

5 Questions 5 Answers 0 Followers

Questions related from Alia Mike

Which buffer to use during buffer exchange?

I need to cleave the trigger factor from my target protein. The current protein is in buffer containing 50mM NaH2PO4H2O, 300mM NaCl, 250mM Imidazole, pH 8. I want to cleave the trigger factor off...

30 November 2021 7,946 2 View

My colony PCR is giving a positive result but my restriction digest doesn't show my inserted gene???

I did a PCR and inserted restriction sites on either side of my gene of interest. I then digested the PCR product and performed ligation into two different plasmids. 5:1, insert vector. I...

30 November 2020 1,642 16 View

After colony PCR I just see long smears...anyone know what this could be?

So I performed transformation on BL21 cells and then used those colonies to perform colony PCR. However results on the gel are just coming up as long smears. I did it twice , with the same...

26 October 2020 6,858 5 View

Is the band that I am seeing after a colony PCR my insert or the entire plasmid itself?

So I have a gene of interest inserted into the pET28a plasmid and I've used it in colony PCR with BL21 cells. The primers I used were pET28 primers. After running the agarose gel with my samples I...

11 October 2020 1,910 3 View

Why can't we do mini prep on BL21 cells?

I want to test to see if my cells do indeed have the plasmid with the insert transformed in them, so I plan to confirm this using a PCR. I've been told we can't do a mini-prep to isolate the...

28 July 2020 6,640 6 View