Anjello Luciano

5 Questions 6 Answers 0 Followers

Questions related from Anjello Luciano

Protein in denaturing purification process-- how to avoid misfolding?

Hello all, I was wondering if I am in danger of crashing out my protein of interest (native fliC) under these conditions: first, I solubilized E. coli cell lysate pellet with 8M urea + 200mM...

10 March 2018 1,480 4 View

How much DEAE sepharose to be added in a column?

Hello, I am pretty new to protein purification and would appreciate some clarification on anion exchange chromatography using GE DEAE Sephacel. 1. Is gravity flow sufficient for this...

10 February 2018 2,110 7 View

Can I substitute Qiagen Buffer QG (from dna extraction kit) for Buffer ERC (Minelute reaction cleanup)?

I want to perform clean up on my digests but currently out of ERC from minElute clean up kit. Can I substitute it with buffer QG ? These are all from qiagen. Somebody told me I could do this but...

05 October 2017 8,297 3 View

DNA purity ratio (260/280) affects ligation ?

Hello, Me and my fellow undergrads are attempting to locate RSVGN insert to pali2 vector. The problem earlier was, we ran out of Qiagen MinElute kit used for clean up after restriction digest. I...

05 October 2017 1,071 2 View

Thoughts on performing PCR on purified, digested dna that has already been amplified (PCR) prior to digestion?

Hello, So I am in the process of ligating RSVGN insert to pali2 vector. I performed PCR for amplification of RSVGN insert. After purifying it, I performed double digestion with Ncol and Spe1...

03 October 2017 4,016 3 View