High throughput sequencing applications have become essential in genome and transcriptome analysis nowadays. Some breast cancer cases are caused due to genetic changes in the BRCA1 and BRCA2 genes. In addition, we validate the use of an absolute standardĬurve based on a dilution series of fluorometrically quantified PCR products.īreast cancer is the most common type of cancer in women worldwide and is the second leading cause of cancer death. We describe here a novel method for creating a reliable standardĬurve using one plasmid containing both alternative transcripts. Primer to quantify isoforms that differ greatly in abundance. We propose the use of a boundary-spanning Overview of three different possibilities for detecting alternative transcripts is given. We use skipping of exon 37 in the NF1 gene as a model to compare and evaluate the different strategies for quantitating splice variants using real-time PCR. To study its application in quantification of splice isoforms. The many advantages of real-time PCR have made this technique attractive ![]() To the intrinsic limitations of conventional methods. So far, accurate quantification of splice variants has been laborious and difficult due A reliable and robust method for measuring the expression of alternatively spliced transcripts is an important step in investigating
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