Patent ID: 6329180
Filing Date: 2001-12-11
Classification: C12Q

Abstract:
A method for detecting a protein altering mutation in a test sequence of a gene comprising:(a) amplifying the test sequence of the gene using a 5' primer, the 5' primer comprising, from its 5' to 3' direction:(i) an RNA polymerase promoter;(ii) a mRNA translation initiation site;(iii) a polynucleotide sequence encoding a peptide tag; and(iv) a polynucleotide sequence that hybridizes to a sequence on the non-coding strand adjacent to the 5' end of the test sequence;(b) transcribing in-vitro the product of step (a);(c) translating in-vitro the product of step (b);(d) purifying the product of step (c) by affinity purification using the peptide tag; and(e) analyzing the purified product of step (d) by mass spectrometry to determine whether a protein altering mutation is present in the test sequence.