Protocols

To assess TP53 mutation status it is recommended to perform gene sequencing of all exons, including the splice junctions and completed with MLPA to detect large deletion/duplications. Immuno-histochemical analysis is not a good method for assessing TP53 mutation status, as it cannot distinguish between the various types of mutations, does not detect truncating mutations (false-negative results), and gives false-positive results (over-expressed WT protein).

• Direct sequencing by Sanger
  
• TP53 sequencing by a next generation sequencing technology (NGS). The detailed list of primers can be downloaded here
  
• MLPA
• Some caution regarding TP53 variants detected by multigene panel testing by NGS: a study by Weitzel et al.,
• SNP sequencing and haplotyping
  

Recent developments on p53 antibodies

• Monoclonal Antibodies against Specific p53 Hotspot Mutants as Potential Tools for Precision Medicine
  
• Understanding p53 functions through p53 antibodies

• Mutalyzer, a package to support checks of sequence variant nomenclature according to the guidelines of the Human Genome Variation Society


• Resources for the systematic analysis of p53 interactions with DNA and other transcription factors.
• p53retriever, for the detection and display of p53 putative response elements (REs) on DNA sequences
• TargetGeneReg, for displaying gene of interest in the p53 and cell cycle gene regulatory networks
• TP53 RNA expression data from TCGA at GEPIA
• Datamed, a search engine for TP53 datasets


• Human Splicing Finder
• Align-AGVGD, for the prediction of the functional impact of missense mutations
• SIFT, for the prediction of the functional impact of missense mutations
• Scorecons, for scoring residue conservation
• The Cancer Genome Interpreter, to identify driver mutations in tumor samples


• LFSPRO, a tool for estimating TP53 Mutation Carrier Probability
• Famdenovo, an algorithm that calculates the probability of a germline mutation to be de novo based on family history data


• Amino-acid letter codes and properties
• Genetic code


• LOVD, a software to build mutation databases