Search found 19 matches

by nukeman
Wed May 30, 2012 9:00 pm
Forum: Molecular Biology
Topic: Idea to determine order of assembly of spliceosome
Replies: 15
Views: 15891

Forgot one thing:

Say I precipitate the samples at different points in time, how would I go on about analysing them? Is it feasible to use MS here or are the differences (when the next snRNP joins in) in snRNP mass enough for me to be able to detect change in mass on a western?
by nukeman
Wed May 30, 2012 6:14 pm
Forum: Molecular Biology
Topic: Idea to determine order of assembly of spliceosome
Replies: 15
Views: 15891

Aha ok. Thank you very much, I just received a mail from my teacher and he actually suggested I do the precipitation, just as you did. From one thing to another though. What is the purpose of a bacteriophage promoter being ligated in to a vector along with your gene? Is it just to ensure that you ge...
by nukeman
Wed May 30, 2012 10:17 am
Forum: Molecular Biology
Topic: Idea to determine order of assembly of spliceosome
Replies: 15
Views: 15891

Isn't it possible to precipitate samples during different times? Like I had planned initially. If the spliceosome assembles stepwise, then wouldn't we be able to detect different masses at different points in time?
by nukeman
Tue May 29, 2012 9:54 pm
Forum: Molecular Biology
Topic: Idea to determine order of assembly of spliceosome
Replies: 15
Views: 15891

Oh and forgive me if I'm asking stupid questions, like I wrote in my initial post, I'm only studying on a basic level.
by nukeman
Tue May 29, 2012 4:44 pm
Forum: Molecular Biology
Topic: Idea to determine order of assembly of spliceosome
Replies: 15
Views: 15891

Re: Idea to determine order of assembly of spliceosome

How would I differentiate my target proteins from the rest of the proteins? Do you mean that I should use DNA/RNA that matches the pre-mRNA and that way isolate the target pre-mRNA along with the associated snRNP:s?
by nukeman
Tue May 29, 2012 12:29 pm
Forum: Molecular Biology
Topic: Idea to determine order of assembly of spliceosome
Replies: 15
Views: 15891

Re: Idea to determine order of assembly of spliceosome

The thing is that the questions are made so that we will pretend that we don't know that much about the spliceosome as we actually do, and from there proceed to map its function and so on. I should have written this more clearly. In essence, that means I "don't know" the binding partners, ...
by nukeman
Mon May 28, 2012 7:48 pm
Forum: Molecular Biology
Topic: Idea to determine order of assembly of spliceosome
Replies: 15
Views: 15891

Idea to determine order of assembly of spliceosome

Hi everyone, I've got this assignment where I'm supposed to present a suggestion of a series of methids to use in order to determine the order of binding of snRNP:s which eventually assemble to a spliceosome. My initial thought was to use in vitro splicing assay with a couple of extra steps. 1. Liga...