Yazen Alnefeesi

4 Questions 8 Answers 0 Followers

Questions related from Yazen Alnefeesi

What should primers be dissolved in for high-fidelity PCR and downstream cloning of amplicons?

I'm amplifying a transgene using Pfu and plan to double-digest it with NotI and XhoI (same buffer) to ligate it into a plasmid. I've never done high-fidelity PCR and I'm not sure what the primers...

25 September 2021 6,640 3 View

How to decide whether a bacterial smear should be fixed with heat or methanol?

I'm curious about the pros and cons of each fixation method, and the kinds of downstream applications that would necessitate the use of one method over the other.

25 September 2021 894 0 View

DNA migrated backwards, and 3 tracking dyes moved as expected. Why did that happen?

Full disclosure: This was the third run using the same gel and running buffer (I added distilled water to compensate for evaporation). I know you're not supposed to reuse running buffer this much,...

09 January 2021 6,694 6 View

Can chloroform be effectively stabilized with isopropanol? And why does ethanol stabilization usually only involve 1% by mass?

The oxidation of chloroform is dangerous because it results in phosgene formation under exposure to UV light. Along with the minimization of UV exposure, this reaction is usually prevented with 1%...

31 December 2020 3,722 3 View