INS 832/13 cells were harvested after transfection with siRNAs or treatment with tunicamycin (final concentration of 2.5 μg/ml), total RNA was isolated using PureLink RNA Mini Kits (Invitrogen no. 12183018A), and cDNA was synthesized using First Strand cDNA Synthesis Kits (OriGene no. NP100042). The following primers were used to amplify spliced and unspliced xbp-1 transcripts: 5′-ATTCTGACGCTGTTGCCTCT-3′ (forward) and 5′-CTCTGGGGAAGGACATTTGA-3′ (reverse).

PCR products were subjected to 2.5% agarose gel electrophoresis. The size difference between products amplified from spliced and unspliced xbp-1 cDNA templates is 26 nucleotides.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.