SALSA MLPA probemix P056-C1 TP53 Lot C1-0215 & lot C1-0214. As compared to version B1 (lot B1-1011) most of the reference and flanking probes have been replaced and several have been added. Furthermore, one probe for TP53 exon 9a and one probe downstream TP53 exon 11 have been added. Moreover, one CHEK2 probe is replaced and one added. Li-Fraumeni syndrome (LFS) is a clinically and genetically heterogeneous inherited cancer syndrome. LFS is characterised by autosomal dominant inheritance and early onset of tumours, multiple tumours within an individual, and multiple affected family members. Approximately 70% of LFS cases and 40% of LFL (Li- Fraumeni like syndrome) cases contain germline mutations in the TP53 gene on chromosomal band 17p13.1. In a subset of non-p53 patients with LFS, mutations have been identified in the CHEK2 gene on chromosomal band 22q12.1. A third LFS locus has been identified on chromosomal band 1q23. The TP53 gene (11 exons) spans ~19 kb of genomic DNA and is located on 17p13.1, about 7,5 Mb from the p-telomere. The P056-C1 TP53 probemix contains probes for each of the 11 exons of the TP53 gene. Three probes are present for the CHEK2 gene on chromosome 22q12.1, one of which is specific for the CHEK2 1100delC mutation. This probe will only give a signal when the mutation is present! Furthermore, flanking probes for both TP53 and CHEK2 are included. Please note that more CHEK2 probes are available in SALSA MLPA probemix P190 CHEK2. No probes for 1q23 LFS locus are present. In addition, 14 reference probes are included in this probemix, detecting several different autosomal chromosomal locations. This P056-C1 TP53 probemix is primarily intended for non-tumour DNA analysis. Nevertheless, the new reference probes target relatively stable chromosomal regions and allow analysis of tumour DNA as well. Please note that Loss of Heterozygosity (LOH) of the TP53 gene is often copy number neutral. LOH without copy number change will not be detected by MLPA. Copy number neutral LOH is indicative of uniparental disomy (UD) and may result in a cell type containing two copies of the TP53 gene that are both inactivated by the same point mutation. SD029 Sample DNA Please note that the mutation-specific probe has only been tested on control plasmids and not on positive human DNA samples with the CHEK2 1100delC point mutation! This SD029 sample DNA is provided with each probemix vial and can be used in data binning in the fragment analysis (see next page). This SALSA probemix is designed to detect deletions/duplications of one or more sequences in the aforementioned genes and to detect the presence of the aforementioned mutation in a DNA sample. Heterozygous deletions of recognition sequences should give a 35-50% reduced relative peak height of the amplification product of that probe. Note that a mutation or polymorphism in the sequence detected by a probe can also cause a reduction in relative peak height, even when not located exactly on the ligation site! In addition, some probe signals are more sensitive to sample purity and small changes in experimental conditions. Therefore, deletions and duplications detected by MLPA should always be confirmed by other methods. Not all deletions and duplications detected by MLPA will be pathogenic; users should always verify the latest scientific literature when interpreting their findings. We have no information on what percentage of defects in these genes is caused by deletions/duplications of complete exons. Finally, note that most defects in this gene are expected to be small (point) mutations which will not be detected by this SALSA test. SALSA probemixes and reagents are sold by for research purposes and to demonstrate the possibilities of the MLPA technique. They are not CE/FDA certified for use in diagnostic procedures. Purchase of the SALSA MLPA test probemixes and reagents includes a limited license to use these products for research purposes. The use of this SALSA MLPA probemix and reagents requires a thermocycler with heated lid and sequence type electrophoresis equipment. Different fluorescent PCR primers are available. The MLPA technique has been first described in Nucleic Acid Research 30, e57 (2002). Related SALSA probemixes P190 CHEK2: Breast cancer susceptibility, genes included: CHEK2, ATM, PTEN, TP53. SALSA P056 TP53 probemix Page 1 of 8
More information Website : www.mlpa.com E-mail : info@mlpa.com (information & technical questions); order@mlpa.com (for orders) Mail : bv; Willem Schoutenstraat 1, 1057 DL Amsterdam, the Netherlands References Vecchio D et al., 2014. Pharmacokinetics, pharmacodynamics and efficacy on pediatric tumors of the glioma radiosensitizer KU60019. Int J Cancer. 136:1445-57. Mitchell G et al., 2013. High frequency of germline TP53 mutations in a prospective adult-onset sarcoma cohort. PLoS One. 8:e69026. Giacomazzi J et al., 2013. Li-Fraumeni and Li-Fraumeni-like syndrome among children diagnosed with pediatric cancer in Southern Brazil. Cancer. 119:4341-9. Rath MG et al., 2013. Prevalence of germline TP53 mutations in HER2+ breast cancer patients. Breast Cancer Res Treat. 139:193 8. Giacomazzi J et al., 2013. TP53 p.r337h is a conditional cancer-predisposing mutation: further evidence from a homozygous patient. BMC Cancer. 13:187. Pinto EM et al., 2012. An identical, complex TP53 mutation arising independently in two unrelated families with diverse cancer profiles: the complexity of interpreting cancer risk in carriers. Oncogenesis. 1:e1. Silva AG et al., 2012. Li-Fraumeni-like syndrome associated with a large BRCA1 intragenic deletion. BMC Cancer. 12:237. Magnusson S et al., 2012. Prevalence of germline TP53 mutations and history of Li-Fraumeni syndrome in families with childhood adrenocortical tumors, choroid plexus tumors, and rhabdomyosarcoma: a population-based survey. Pediatr Blood Cancer. 59:846-53. Magnusson S. et al., 2012. Increased incidence of childhood, prostate and breast cancers in relatives of childhood cancer patients. Fam Cancer. 11:145-55. Ruijs MW et al., 2010. TP53 germline mutation testing in 180 families suspected of Li-Fraumeni syndrome: mutation detection rate and relative frequency of cancers in different familial phenotypes. J Med Genet. 47:421-8. Mouchawar J et al., 2010. Population-based estimate of the contribution of TP53 mutations to subgroups of early-onset breast cancer: Australian breast cancer family study. Cancer Res. 70:4795-800. Pinto C et al., 2009. TP53 germline mutations in Portugal and genetic modifiers of age at cancer onset. Fam Cancer. 8:383-90. Interpretation of the results - Data analysis The P056-C1 TP53 probemix contains 43 MLPA probes with amplification products between 128 and 489 nt. This includes one probe, specific for the CHEK2 1100delC mutation which will only generate a signal when the mutation is present. In addition, it contains 9 control fragments generating an amplification product smaller than 120 nt: four DNA Quantity fragments (Q-fragments) at 64-70-76-82 nt, three DNA denaturation control fragments (D-fragments) at 88-92-96 nt, one X-fragment at 100 nt and one Y-fragment at 105 nt. More information on how to interpret observations on these control fragments can be found in the MLPA protocol. SD029 Sample DNA The SD029 Sample DNA provided with this probemix can be used as Binning DNA sample for binning of the CHEK2 1100delC mutation-specific probe (CHEK2 1100delC probe 18318-L26751). Inclusion of one reaction with SD029 DNA in MLPA experiments is recommended as it can be used to aid in data binning of the peak pattern using Coffalyser.NET software. Binning SD should never be used as a reference sample in the MLPA data analysis. Neither should it be used in quantification of mutation signal(s), as for this purpose true mutation/snp positive patient samples or cell lines should be used. For further details, please consult the SD029 Binning DNA product description provided (also available online: www.mlpa.com). This product is for research use only. Data generated by this probemix can first be normalised intra-sample by dividing the peak height of each probe s amplification product by the total peak height of only the reference probes in this probemix (block normalisation). Secondly, inter-sample normalisation can be achieved by dividing the intra-normalised probe ratio in a sample by the average intra-normalised probe ratio of all reference samples. Please, note that this SALSA P056 TP53 probemix Page 2 of 8
type of normalisation assumes no changes occurred in the genomic regions recognised by the reference probes. Data normalisation should be performed within one experiment. Only samples purified by the same method should be compared. Confirmation of most exons deletions and amplifications can be done by e.g. Southern blotting, long range PCR, qpcr, FISH. Note that Coffalyser, the MLPA analysis tool developed at, can be downloaded free of charge from our website www.mlpa.com. CHEK2 1100delC probe This P056 probemix contains a CHEK2 1100delC mutation-specific probe. Results obtained with this CHEK2 1100delC probe should be treated with caution! We have received reports of experiments at three different laboratories in which the CHEK2 1100delC peak in the P045 BRCA2 probemix spuriously appeared in known normal controls as well as in all other samples. We have observed the same problem with the similar CHEK2 probe in the P190 CHEK2 probemix. The cause of this phenomenon is unknown and despite several attempts, we were not able to reproduce the results. In case the peak of the 1100delC probe is observed at 208 nt, always confirm the result by sequencing. Many copy number alterations in healthy individuals are described in the database of genomic variants: http://dgv.tcag.ca/dgv/app/home. For example, a duplication of a complete gene might not be pathogenic, while a partial duplication or a deletion may result in disease. For some genes, certain in-frame deletions may result in a very mild, or no disease. Copy number changes of reference probes are unlikely to be the cause of the condition tested for. Users should always verify the latest scientific literature when interpreting their findings. Warning: In case the P056-C1 probemix is used on tumour samples it should be considered that MLPA tumour analysis provides information on the average situation in the cells from which the DNA sample was purified. Gains or losses of genomic regions or genes may not be detected if the percentage of tumour cells is low. Furthermore, although reference probes are located in stable regions that are not frequently altered in copy number in various cancer types, there is always a possibility that one or more reference probes do show a copy number alteration in a sample. Normal copy number variation in healthy individuals is described in the database of genomic variants: http://dgv.tcag.ca/dgv/app/home. When in doubt, users should always verify the latest updates of the database and scientific literature when interpreting their findings. This probemix was developed at. Info/remarks/suggestions for improvement: info@mlpa.com. SALSA P056 TP53 probemix Page 3 of 8
Table 1. SALSA MLPA P056-C1 TP53 probemix Length Chromosomal position SALSA MLPA probe (nt) reference TP53 CHEK2 64-70-76-82 Q-fragments: DNA quantity; only visible with less than 100 ng sample DNA 88-92-96 D-fragments: Low signal of 88 or 96 nt fragment indicates incomplete denaturation 100 X-fragment: Specific for the X chromosome 105 Y-fragment: Specific for the Y chromosome 128 * Reference probe 19616-L26684 4p13 135 Reference probe 16316-L21434 3q21.3 140 EFNB3 probe 03962-L21069 17p13.1 147 * MPDU1 probe 19643-L26685 17p13.1 153 EFNB3 probe 01984-L21070 17p13.1 160 Reference probe 09787-L10202 15q21.1 166 TP53 probe 01588-L06028 exon 1 175 * TP53 probe 19637-L26296 exon 11 184 * POLR2A probe 19647-L10408 17p13.1 193 * Reference probe 11556-L26606 5q31.2 199 TP53 probe 01996-L26321 exon 2a 208 CHEK2 probe 18318-L26751 1100delC 216 ± TP53 probe 02375-L26750 exon 2d 224 * + TP53 probe 19638-L26297 exon 9a 230 * Reference probe 17130-L26574 11p11.2 238 * «AKAP10 probe 19648-L00940 17p11.2 247 * Reference probe 08728-L08739 9q21.13 255 TP53 probe 02376-L21073 exon 4b 265 HSCB probe 06800-L02040 22q12.1 274 * Reference probe 17450-L21206 16p13.2 283 ± TP53 probe 01999-L21074 exon 7 300 TP53 probe 17420-L21142 exon 3 310 Reference probe 07028-L06639 14q11.2 318 TP53 probe 17421-L21315 exon 5 328 * Reference probe 13397-L26608 6q12 335 * ATP1B2 probe 19884-L26749 17p13.1 346 TP53 probe 17422-L21144 exon 10 355 * «CHEK2 probe 19654-L26320 exon 13 364 * «NF2 probe 19698-L26319 22q12.2 372 * Reference probe 14835-L26609 1p34.1 382 * ATP1B2 probe 19645-L26316 17p13.1 391 TP53 probe 17423-L21145 exon 8 401 * TP53 probe 19650-L21141 exon 6 409 TP53 probe 02263-L01749 exon 1 420 * Reference probe 08839-L08899 2p13.2 427 * CHEK2 probe 06631-L06189 exon 8 436 * MPDU1 probe 19646-L26317 17p13.1 445 TP53 probe 17424-L21146 exon 11 454 TP53 probe 17425-L21147 exon 4b 463 * POLR2A probe 09951-L26312 17p13.1 471 Reference probe 00979-L21316 10p14 481 * Reference probe 14203-L15817 2q12.3 489 * Reference probe 19137-L25693 21q22.2 * New in version C1 (from lot C1-0214 onwards). Changed in version C1 (from lot C1-0214 onwards). Small change in length, no change in sequence detected. Mutation-specific probe. This probe will only generate a signal when the CHEK2 1100delC mutation is present. It has been tested on artificial test DNA but not on positive human samples! Results obtained with this CHEK2 1100delC probe should be treated with caution! Please, read the note on page 3. Flanking probe. Included to facilitate the determination of the extent of a deletion/duplication. Copy number alterations of flanking and reference probes are unlikely to be related to the condition tested. «This probe is located within, or close to, a very strong CpG island. A low signal of this probe can be due to incomplete sample DNA denaturation, e.g. due to the presence of salt in the sample DNA. SALSA P056 TP53 probemix Page 4 of 8
± SNP rs201753350 could influence the probe signal at 216 nt and SNP rs121912660 could influence the probe signal at 283 nt. In case of apparent deletions, it is recommended to sequence the region targeted by this probe. + Exon numbering for exon 9a is indicated according to NCBI sequence NM_001126114.2 and NG_017013.2. Exon 9a is present in some alternative transcript variants only. Ligation site of this probe is located on a common mutational hotspot both in germline and somatic samples as reported by IARC TP53 Database (http://p53.iarc.fr/). In case of apparent deletions, it is recommended to sequence the region targeted by this probe. Table 2. P056 probes arranged according to chromosomal location Table 2a. Reference probes. Length SALSA MLPA Gene (nt) probe Location Partial sequence MV (24 nt adjacent to ligation site) location 372 14835-L26609 UROD 1p34.1 AAGCACCATGGC-TCAGGCCAAGCG 01-045.252174 420 08839-L08899 DYSF 2p13.2 TGCCATGAAGCT-GGTGAAGCCCTT 02-071.767082 481 14203-L15817 EDAR 2q12.3 AAAGCCCACCAA-GAGGTATGTGGA 02-108.893314 135 16316-L21434 RAB7A 3q21.3 CACAATAGGAGC-TGACTTTCTGAC 03-129.999512 128 19616-L26684 ATP8A1 4p13 CAGATTCTTCTT-CGAGGAGCTCAG 04-042.278439 193 11556-L26606 MYOT 5q31.2 CAGATCTCGGCT-ATACCTCCACCA 05-137.249703 328 13397-L26608 EYS 6q12 ATGGTAAGATTA-ACTGAACCCTCT 06-064.573993 247 08728-L08739 PCSK5 9q21.13 GACTATGAAGAA-TGTGTCCCTTGT 09-078.045346 471 00979-L21316 UPF2 10p14 TGCCATTCCTTT-GCATCTCAAAAG 10-012.018627 230 17130-L26574 MYBPC3 11p11.2 GTGCCTCAGTGA-CCAGGCTGGCTC 11-047.309963 310 07028-L06639 RPGRIP1 14q11.2 GAGGTTCCCATT-GAAGCTGGCCAG 14-020.866425 160 09787-L10202 SPG11 15q21.1 GGGACACATTCA-GGACTCAACAGA 15-042.674827 274 17450-L21206 GRIN2A 16p13.2 TGCAGGATTATA-ATCTCACAATCT 16-009.761070 489 19137-L25693 PSMG1 21q22.2 TGGAAGCTTTTA-AGCCTATACTTT 21-039.471231 Table 2b. Chromosome 17p probes. Length SALSA MLPA Gene / Exon (nt) probe Location / Ligation site Partial sequence (24 nt adjacent to ligation site) Distance to next probe centromeric 238 19648-L00940 AKAP10, ex 3 17p11.2 AGGACCAAGTCA-TGTTGCAATCAA 12254.7 kb 140 03962-L21069 EFNB3, ex 2 17p13.1 TCTCCTAATTAT-GAGTTCTACAAG 2.5 kb 153 01984-L21070 EFNB3, ex 1 17p13.1 GTCCGGGCTGAA-GAGCCAGGCAGC 17.9 kb TP53, at 17p13.1. Ligation sites are indicated according to NM_000546.5, except for exon 9a, which is indicated according to NM_001126114.2 (+) (transcript variant 3, also known as p53beta). Exon numbering is indicated according to the reference sequences NM_000546.5 and NG_017013.2. start codon 203-205 (ex 2a) 409 02263-L01749 exon 1 67 nt before ex 1 CTTCCTCCGGCA-GGCGGATTACTT 0.2 kb 166 01588-L06028 exon 1 118-119 TCCGGGGACACT-TTGCGTTCGGGC 10.8 kb 199 01996-L26321 exon 2a 177-178 CTCTTGCAGCAG-CCAGACTGCCTT 0.2 kb 216 ± 02375-L26750 exon 2d 290-291 TTCCTGAAAACA-ACGTTCTGGTAA 0.3 kb 300 17420-L21142 exon 3 511-510, reverse TAGCTGCCCTGG-TAGGTTTTCTGG 0.8 kb 255 02376-L21073 exon 4b 606-607 CAAGATGTTTTG-CCAACTGGCCAA 0.1 kb 454 17425-L21147 exon 4b 696-697 CATCTACAAGCA-GTCACAGCACAT 0.2 kb 318 17421-L21315 exon 5 795-796 TATCCGAGTGGA-AGGAAATTTGCG 0.7 kb 401 19650-L21141 exon 6 891-892 CTCTGACTGTAC-CACCATCCACTA 0.5 kb 283 ± 01999-L21074 exon 7 1041-1042 CTGTCCTGGGAG-AGACCGGCGCAC 0.2 kb 391 17423-L21145 exon 8 1155-1156 CTCTCCCCAGCC-AAAGAAGAAACC 0.2 kb 224 + 19638-L26297 exon 9a 1196-1195, reverse GCTGGTCTGGTC-CTTTAAAATATA 2.7 kb 346 17422-L21144 exon 10 1248-1249 TGAGGCCTTGGA-ACTCAAGGATGC 1.0 kb 445 17424-L21146 exon 11 1360-1361 CTCATGTTCAAG-ACAGAAGGGCCT 3.3 kb stop codon 1382-1384 (ex 11) 175 19637-L26296 downstream 2091 nt after ex 11, TGAAGCCATGAG-GAAATTGGGAGA reverse 9.8 kb 382 19645-L26316 ATP1B2, ex 7 17p13.1 AGAACCACCTTG-TCCTCAATTACA 5.0 kb 335 19884-L26749 ATP1B2, ex 1 17p13.1 CCGCGCCACCAA-GATGGTCATCCA 63.8 kb SALSA P056 TP53 probemix Page 5 of 8
436 19646-L26317 MPDU1, ex 7 17p13.1 TTTTCTTTTAGT-GGAAACAAATGG 1.7 kb 147 19643-L26685 MPDU1, ex 3a 17p13.1 GCTGCCCCAGGT-GTTTAAAATCCT 87.8 kb 463 09951-L26312 POLR2A, ex 8 17p13.1 CGCCAAGTACAT-CATCCGAGACAA 1.8 kb 184 19647-L10408 POLR2A, ex 3 17p13.1 GAAGACAATGAA-AGTTTTGCGCTG - telomeric + Ligation site and exon numbering for exon 9a is indicated according to NCBI sequence NM_001126114.2 and NG_017013.2. Exon 9a is present in some alternative transcript variants only. Ligation site of this probe is located on a common mutational hotspot both in germline and somatic samples as reported by IARC TP53 Database (http://p53.iarc.fr/). In case of apparent deletions, it is recommended to sequence the region targeted by this probe. ± SNP rs201753350 could influence the probe signal at 216 nt and SNP rs121912660 could influence the probe signal at 283 nt. In case of apparent deletions, it is recommended to sequence the region targeted by this probe. Flanking probe. Included to facilitate the determination of the extent of a deletion/duplication. Copy number alterations of flanking and reference probes are unlikely to be related to the condition tested. Table 2c. Chromosome 22 probes. Length SALSA MLPA Gene (nt) probe Location/ Ligation site Partial sequence (24 nt adjacent to ligation site) Distance to next probe CHEK2, at 22q12.1. Ligation sites and exon numbering are indicated according to NM_007194.3 which represents the predominant transcript and encodes isoform a. telomeric 364 19698-L26319 NF2, ex 4 22q12.2 CTCCTGAGGCTT-CTGTGCTCCTGG 898.3 kb 265 06800-L02040 HSCB, ex 2 22q12.1 TGGCTGAAGAAA-TCTGGGTGGACA 40.4 kb 427 06631-L06189 CHEK2, ex 8 937-938 TCATCAAGATTA-AAAACTTTTTTG 7.7 kb 208 18318-L26751 CHEK2, ex 11 1100delC 1173-1171, reverse TGCCCAAAATCA-TAATCTAAAATT 1.7 kb 355 19654-L26320 CHEK2, ex 13 54 nt before ex 13 TCTGGCATACTC-TTACTGATAATA - centromeric Flanking probe. Included to facilitate the determination of the extent of a deletion/duplication. Copy number alterations of flanking and reference probes are unlikely to be related to the condition tested. Mutation-specific probe. This probe will only generate a signal when the CHEK2 1100delC mutation is present. The current 1100delC mutation specific probe cannot distinguish between the 1100delC sequence change present in the CHEK2 gene or its pseudogene. In case a positive signal is obtained with this probe, Sanger sequencing can clarify whether the mutation is present in the actual gene. Note: We have adopted the exon numbering mentioned in the Genbank NM_000546.5 transcript variant 1 and NG_017013.2, which are reference standards in the RefSeqGene project. This exon numbering might be different as compared to (older) literature! One probe is included for exon 9a, which is only present in some alternative transcripts encoding the p53beta, delta40p53beta, delta133p53beta and delta160p53alpha variant proteins. Complete probe sequences are available on request: info@mlpa.com. Please, notify us of any mistakes: info@mlpa.com. SALSA P056 TP53 probemix Page 6 of 8
SALSA MLPA probemix P056-C1 TP53 sample pictures Figure 1. Capillary electrophoresis pattern of a sample of approximately 50 ng human male control DNA analysed with SALSA MLPA probemix P056-C1 TP53 (lot C1-0215). Figure 2. Capillary electrophoresis pattern of SD029 sample DNA (approximately 50 ng) analysed with SALSA MLPA probemix P056-C1 TP53 (lot C1-0215). The location of the CHEK2 1100delC mutation-specific probe at 208 nt is indicated with an arrow. SALSA P056 TP53 probemix Page 7 of 8
Implemented Changes compared to the previous product description versions. Version 16 23 April 2015 (54) - Product description adapted to a new lot (lot number added, small changes in Table 1 and Table 2, new pictures included). - Information added about the reference standard NG_017013.2, which is used for exon numbering. - Various minor textual changes on the whole document. - Electropherogram of the female reaction removed from page 7. - SD information adjusted on page 2. Version 15 02 February 2015 (54) - Information about 1100delC mutation specific probe is added. Version 14 (53) - Reference added on page 2. - Information about two probes (255 and 283 nt) added in Table 1 and Table 2. Version 13 (53) - Product description adapted to a new lot (lot number added, small changes in Table 1 and Table 2, new pictures included) - A note about exon numbering added below Table 1. - Various minor textual changes on the whole document. Version 12 (48) - Five new references added on page 2. Version 11 (48) - Electropherogram pictures using the new MLPA buffer (introduced in December 2012) added. Version 10 (48) - Exon numbering and ligation sites changed for TP53 in tables and note on page 4 modified accordingly. Version 9 (48) - Product description adapted to a new lot (lot number added, small changes in Table 1 and Table 2, new pictures included). Small changes of probe lengths in Table 1 and 2 in order to better reflect the true lengths of the amplification products. - References added on page 2. - Various minor textual changes on the whole document. - Various minor layout changes. - Data analysis method has been modified. - Sentence when only small numbers of samples are tested, visual comparison of peak profiles should be sufficient removed from data analysis section. - Additional information provided on page 2 about the mutation-specific probe and control plasmid. - Warning added on page 2, in case this probemix is being used for tumour DNA analysis. - NM_001126118.1 have been used for ligation site indication and exon numbering, therefore some differences have been generated as compared to the older nomenclature in table 1 and table 2. Version 8 (45) - Product description adapted to a new lot (lot number added, small changes in Table 1 and Table 2, new picture included). - Small changes of probe lengths in Table 1 and 2 in order to better reflect the true lengths of the amplification products. - Warning added on page 1 on the use of this product to confirm LOH results in tumour DNA. - Warning added below Table 2 that the exon numbering used is new. SALSA P056 TP53 probemix Page 8 of 8