I have a plasmid with an I-SceI restriction site which I want to only cut once, which I do not believe can happen with an I-SceI expression vector.
I am currently looking for a near diploid human cancer cell line, such as HCT116 and U87-MG. Preferably XY with modal chromosome # between 40-50, "normal" DNA repair pathways, and able to easily...
06 July 2015 4,695 2 View
Hello All, I am currently struggling with getting a Gibson Assembly to work. I am trying to insert a 2.5 Kb insert into a roughly 12 Kb plasmid (modified pLEX-MCS). I have had the G.A work in the...
11 December 2014 4,283 7 View
Hello All, I am trying to do a simple fluorescence plus Giemsa stain/acridine orange differential stain for chromosomes. I have had great success in the past using MEF/CHO/and various cancer cell...
11 December 2014 898 2 View
Afternoon everyone, I am currently using Q5 polymerase from NEB to add restriction sites to an insert in a pBabepuro plasmid (~5400 Kb vector and 2.2 Kb insert). I have designed the primers to...
09 October 2014 4,545 4 View
Hello, Addgene does not have the complete sequence published. Also, does anyone know if myc is truncated on both the N and C terminus, a primer designed to target the start of the myc CDS,...
08 September 2014 1,467 1 View
I have been running several western blots over the past few days and have had the same issue where large proteins seem to stain nicely where as the smaller (~50 Kda) stain very poorly. I am using...
05 June 2013 9,776 4 View
I'm interested in learning about enzyme usage. Could someone explain the basics of working with enzymes? I understand they're different from standard chemicals, but I'm uncertain about the...
01 August 2024 5,460 5 View
I want to choose a resarch topic regarding enzyme inhibition. So I did my research and found out most of the diseases that originate from enzymes were actually caused by the "deficiency" of...
30 July 2024 2,483 5 View
I am working on carbonic anhydrase immobilization into MOFs. I am facing problems with low enzyme loading.. The other issue is that when using p-NPA activity test to detect the activity of the...
20 July 2024 1,440 3 View
Good morning, I am trying to produce a recombinant protein. Unfortunately I one of the two restriction enzymes that I use do not cut at the end of the MCS. After the digestion 15 nucleotides of...
17 July 2024 2,787 4 View
Is it appropriate to place the ATG codon in front of the gene of interest, since there is a secretion signal that has its own ATG in front of this gene? I need my protein of interest to be...
10 July 2024 3,050 5 View
كيف يساهم انزيم الكاتلاز في عملية التمثيل الضوئي في النباتات؟
09 July 2024 7,512 1 View
ماهو دور انزيم الكاتليز في النباتات؟
09 July 2024 5,824 2 View
what is the protocol for enzyme freeze drying?
09 July 2024 6,464 6 View
Hi We have alpha-amylase enzyme that written 5 units /mg protein on its container. How do I make 10 cc of enzyme solution with a concentration of 0.2 units/mL?
08 July 2024 2,285 2 View
Several times by placing a restriction on the upper part of the membrane with the aim of preventing the drug from being added from this side, we tried to keep the drug at the top of the leaf, but...
29 June 2024 9,688 1 View