[Arabidopsis] Questions regarding a big gene

[wang zhibib]

Hi Everyone,

I am wondering if anyone could help me the following
questions.  I am working on a big gene more than 10kb.
 
1. I had problem with RNA blotting. Any tip to
transfer total RNA well to filter;
2. Protein sequence analyses identified three NLS
(nuclear localization signal) and dispersed in gene. 
Do I need absolutely include three NLS to determine
its subcellular localization? It took too much time
for me to get the full length cDNA. Will it be
sufficient to use one or two NLS to fuse GFP?

Thank you in advance.

Zhibin

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