Search found 4 matches

by polkadotshorts
Mon Dec 10, 2012 7:12 pm
Forum: Genetics
Topic: How to clone a gene with lots of splicing intermediates?
Replies: 6
Views: 6330

Re:

that would suggest there is some small alternative-intron in your exon, which can be thus split why are you using primers only for one exon at a time? Is your aim to clone all variants or are you fine with only some? Well, we initially had 4 sets of primers. I was going to amplify 4 overlapping reg...
by polkadotshorts
Sat Dec 08, 2012 3:52 pm
Forum: Genetics
Topic: How to clone a gene with lots of splicing intermediates?
Replies: 6
Views: 6330

Re:

Hardly will someone answer in half hour. Where do you want to express it? In what system? If you wanted to use eukaryotic system (yeast or best plants), you could clone genomic version including the introns and let the cell to solve your problem. Hello, thank you for your response sorry, this is my...
by polkadotshorts
Sat Nov 24, 2012 7:18 pm
Forum: Genetics
Topic: How to clone a gene with lots of splicing intermediates?
Replies: 6
Views: 6330

Anyone?
by polkadotshorts
Sat Nov 24, 2012 6:49 pm
Forum: Genetics
Topic: How to clone a gene with lots of splicing intermediates?
Replies: 6
Views: 6330

How to clone a gene with lots of splicing intermediates?

I have been trying very very hard to clone the ligase 6 gene in Arabidopsis overexpress it. However, it has been proving very difficult due to the high amount of splicing intermediates. I have so far used varying primers on the cDNA of arabidopsis in order to get some fragments that I could then lig...