A similar pattern of habitat-driven taxonomic bias was seen in the first ecogenomic survey of sequenced microbial genomes, whereby 67% of the buy inhibitor sequenced marine microbes were phototrophs [8]. Genome Pairs The study of phages and hosts intrinsically lends itself to taking advantage of what Martiny and Field describe as “one of the most exciting and underutilized aspects of the genome collection” [8]: genome pairs. A genome pair occurs when organisms with potential natural interactions are both sequenced, e.g., a phage and host. These associations have revealed patterns in genome biology, such as how well pairs correlate based on %G+C content or tetranucleotide genome signatures [8,32]. Such pairs can (and soon will) rapidly evolve to complex networks as multiple phages infecting the same host, or multiple hosts infected by the same phage, are sequenced.

This complexity obviates the need for the basic units, the pairs, to be explicitly documented (as called for by MIGS) in a structured form. This is possible through the GCDML ‘original host’ and ‘alternate host’ fields, where they can be stored for automated retrieval and network visualization. This process was just barely possible by hand with the 27 marine phage genomes, and reveals interesting trends (Figure 7). Thus far, most cyanophage-cyanobacteria associations are one-to-one pairs, though many cyanophages are known with broad host ranges [33]. Furthermore, such visualization leads to hypotheses about the ‘lone phages’, such as Phage phiJL001, Halomonas phage HAP-1, and Cyanophage Syn5, which lack a sequenced host, but which exist in phylogenetic groups with related sequenced hosts (Figure 7).

The current map is useful in designing future sequencing ventures to answer targeted questions, such as “What drives phage host range�� and ��what are the genomic consequences of all members belonging to the same network?” Figure 7 Network of ‘genome pairs’ and interactions between sequenced marine phages and sequenced hosts. Solid lines link phages (empty circles) to the host strain (solid circles) they infect; dashed lines connect phages to the host species (but not necessarily … Environmental parameters Additionally, the 27 ��mappable�� genomes can be further analyzed in their environmental context using emerging resources, such as megx.

net, to (i) ��put them on the map�� (Panel a of Figure 4; [14]), and (ii) extract interpolated environmental data, though only possible for the eight genomes Brefeldin_A where depth is reported and which are not too close to the coast (Table 1). Preliminary analysis of the megx.net interpolated data available in the GCDML reports revealed that, based on physical-chemical parameters across sample sites, e.g., the four phages isolated from the Sargasso Sea cluster together, while Cyanophage PSS2 appears to be an outlier (Panel b of Figure 4).

arsenaticum. Within this genome, 145 non-coding RNA genes are described. These include a single operon encoding 16S and 23S ribosomal RNA, the associated 5S rRNA, the 7S signal recognition particle(SRP), and the RNase P RNA. Ganetespib cost There are 47 annotated tRNA genes, plus a single tRNA pseudogene. Also included are 83 predicted C/D box sRNA genes and nine additional H/ACA-like sRNA, each of which has been transcriptionally validated [31]. The non-coding RNA content of the P. oguniense genome has become the most extensively annotated among crenarchaeal genomes to date. The use of a not-quite-clonal cell population for DNA isolation, coupled with ultra-deep sequencing has provided a view of three major inversions that are each present in over 17% of the sample population.

The boundaries of one of these inversions are defined by an inverted repeat encoding a duplication of glutamate dehydrogenase (GluDH). Notably, this duplication appears to be present in each of the currently sequenced Pyrobaculum members, suggesting that those genomes may also host similar inversions. A second inversion has at its termini another inverted duplication, encoding a gene associated with one of the paREP members and a CRISPR-associated gene. It remains unclear if these common structural variants impart a physiological advantage, and if so, how the variation provides utility to its host. Based on our expanded genome diversity observations, we suggest that avoiding the use of a strictly clonal population for sequencing purposes can provide a significant benefit to understanding both the biology of the host and a clearer understanding of the genome dynamics of the species.

Acknowledgements Sequencing was provided by the UCSC Genome Sequencing Center. We would like to thank Nathan Boyd, Eveline Hesson and Nader Pourmand for their expertise and advice in this work. This work was supported by National Science Foundation Grant DBI-0641061 (T.L. and D.B.) and the Graduate Research and Education in Adaptive Bio-Technology (GREAT) Training Program sponsored by the University of California Biotechnology Research and Education Program (D.B.).
Alistipes senegalensis strain JC50T (= CSUR P156= DSM 25460) is the type strain of A. senegalensis sp. nov. This bacterium was isolated from the stool of a healthy Senegalese patient as part of a ��culturomics�� study aiming at cultivating all species in human feces, individually.

Bacterial species definition is a matter of debate. This is notably due to the high cost, poor reproducibility and inter-laboratory comparability Batimastat of the ��gold standard�� of DNA-DNA hybridization and G+C content determination [1]. In contrast, the development of PCR and sequencing methods, both of which are now widely available and cost-effective, has profoundly changed the way of classifying prokaryotes.

Article downloads on the SIGS and PMC sites map to 15,350 unique IP addresses located in 4,377 cities in 152 countries. Although SIGS has not been publishing long enough to estimate our impact factor, 93 articles have been cited a total of 271 times in articles included in the Cite-by-Linking program of Cross-Ref. Moving forward Our experience with the template AZD-2281 for Short Genome Reports has been largely successful. The layout of content is highly predictable and simplifies writing, reviewing, editing and reading these articles. Yet, we are exploring the possibility of some minor changes to the tabular layout in 2012, to accommodate an anticipated influx of articles from the Thousand Genome Project, which represents the second phase of the GEBA initiative.

Although it is unlikely that we will be able to ��auto-generate�� manuscripts as a part of the sequencing and annotation pipeline, this represents an early attempt to capture and standardize much of the summarized data that is incorporated into Short Genome Reports. This will also give us an opportunity to explore how to more tightly integrate the literature and databases. The second major change for 2012 deals with funding SIGS in the future. We were very fortunate in that seed funding for SIGS was provided through grants from the Office of the Vice President of Research and Graduate Studies of Michigan State University and the Office of Biological and Environmental Research of the US Department of Energy. This has provided us with the opportunity to underwrite the publication costs of articles appearing in Volumes 1 �C 4 and a limited number of articles in Volume 5.

However, like other open access publications we need to institute a cost recovery mechanism to sustain publication of SIGS. More information about the publication fees is included in the Instructions to Authors.
A representative genomic 16S rRNA sequence of D. thermolithotrophum BSAT was compared using NCBI BLAST [3,4] under default settings (e.g. considering only the high-scoring segment pairs (HSPs) from the best 250 hits) with the most recent release of the Greengenes database [5] and the relative frequencies of taxa and keywords (reduced to their stem [6] were determined, weighted by BLAST scores. The most frequently occurring genera were Desulfurobacterium (30.3%), Thermoanaerobacter (18.8%), Thermovibrio (14.2%), Balnearium (11.0%) and Persephonella (4.1%) (80 hits in total). Regarding the two hits to sequences from members of the species, the average identity within HSPs was 98.9%, whereas the average coverage by HSPs was 92.8%. Regarding the single hit to sequences from other GSK-3 members of the genus, the average identity within HSPs was 98.6%, whereas the average coverage by HSPs was 64.4%.

2% and an HSP coverage of 99.9%. The most frequently occurring keywords within the labels of all environmental selleck chemicals Cabozantinib samples which yielded hits were ‘marine’ (3.0%), ‘lake’ (2.9%), ‘depth’ (2.7%), ‘water’ (2.6%) and ‘zone’ (2.5%) (249 hits in total) and corresponded with the habitat from which strain UST20020801T was isolated. Figure 1 shows the phylogenetic neighborhood of O. hongkongensis in a 16S rRNA based tree. The sequences of the two identical 16S rRNA gene copies in the genome do not differ from the previously published 16S rRNA sequence (“type”:”entrez-nucleotide”,”attrs”:”text”:”AB125062″,”term_id”:”38093231″,”term_text”:”AB125062″AB125062). Figure 1 Phylogenetic tree highlighting the position of O. hongkongensis relative to the type strains of the other species within the family Cryomorphaceae.

The tree was inferred from 1,409 aligned characters [21,22] of the 16S rRNA gene sequence under the maximum … Table 1 Classification and general features of O. hongkongensis UST20020801T according to the MIGS recommendations [29] and NamesforLife [30]. O. hongkongensis UST20020801T is a Gram-negative, halophilic, non-flagellated, motile, and rod-shaped bacterium (Figure 2) [1]. Colonies are orange, convex, smooth, glistening and translucent with an entire margin [1]. Cells are 0.3-0.5 ��m in width and 0.5-4.0 ��m in length [1]. The strain does not sporulate [1]. Cells are strictly aerobic heterotrophs requiring Na+, Mg2+, sea salts and yeast extract or peptone for growth [1]. Growth occurs between 4��C and 37��C with an optimum at 25��C-33��C [1]. The pH range for growth is 5.

2-9.0 with an optimum at pH 6.0-8.0 [1]. The salinity range for growth is 1.0-7.5% NaCl as well as 15-100% sea-water [1]. Yeast extract, peptone or starch is required for growth [1]. Ampicillin (10 ��g), chloramphenicol (30 ��g), erythromycin (10 ��g), penicillin G (2U), rifampicin (10 ��g), streptomycin (10 ��g), tetracycline (30 ��g) and polymyxin B (300 U) inhibited Cilengitide growth whereas cells were resistant to kanamycin (10 ��g), gentamycin sulphate (10 ��g) and spectinomycin (10 ��g) [1]. Cells contain oxidase, catalase and alkaline phosphatase [1]. Figure 2 Scanning electron micrograph of O. hongkongensis UST20020801T Chemotaxonomy The fatty-acid profile of strain UST20020801T differs significantly from those of other members of the Cryomorphaceae [1]. The principal cellular fatty acids of strain UST20020801T were the following saturated branched-chain fatty acids: iso-C15:0 G (28.0%), iso-C15:0 (18.7%), iso-C17:0 3-OH (18.1%), iso-C17:1 ��9c (7.3%), iso-C15:0 3-OH (4.9%), and a summed feature containing iso-C15:0 2-OH and/or C16:1��7c (10.0%) [1].

enterica serovar Typhi P-stx-12, S. enterica serovar Typhi CT18, and S. Ganetespib buy enterica serovar Typhi Ty2 CRISPR Region By using the CRISPR Finder tool, one CRISPR repeat region with a length of 394 bp was identified in the S. enterica serovar Typhi P-stx-12 genome. The CRISPR region starts at the position 2,900,675 and ends at the position 2,901,069 with 6 spacers in between. The confirmed CRISPR has the following direct repeat consensus sequence: CGGTTTATCCCCGCTGGCGCGGGGAACAC. Strains CT18 and Ty2 also have a single CRISPR repeat region with the lengths of 385 bp and 394 bp, respectively. The location for the CRISPR region of all three strains falls within the region of 2.9 Mbp on the chromosome. All the strains have 6 spacers and share the common direct repeat consensus sequence.

It is worth noting that the CRISPR region, including the length and the spacer sequence, of S. enterica serovar Typhi P-stx-12 is exactly identical to S. enterica serovar Typhi Ty2. It suggests a strong evidence of their evolutionary relevance and shows that the CRISPR region in S. enterica serovar Typhi is conserved. As CRISPRs function as a prokaryotic immune system and confer resistance towards plasmids and phages (thus interfering with the spread of antibiotic resistance and virulence factors), it is reasonable to find only one CRISPR with very few spacers in this pathogen as compared to other bacterial strains that are not pathogenic [49]. Acknowledgement This work was supported by APEX funding (Malaysia Ministry of Higher Education) to the Centre for Chemical Biology, Universiti Sains Malaysia; and the contributions of the Council of Scientific and Industrial Research (CSIR), New Delhi, India.

Notes Abbreviations: NCBI- National Center for Biotechnology Information RDP- Ribosomal Database Project
Strain YIM 70093T (= DSM 44683T) is the type strain of the species Corynebacterium halotolerans [1] and was originally isolated from saline soil in Xinjiang Province in western China. The genus Corynebacterium is comprised of Gram-positive bacteria with a high G+C content. It currently contains over 80 members [2] isolated from diverse backgrounds like human clinical samples [3] and animals [4], but also from soil [5] and ripening cheese [6]. Within this diverse genus, C. halotolerans has been proposed to form a subclade together with C. freneyi and C. xerosis [1].

Data concerning salt tolerance is not available for most corynebacteria, but C. halotolerans YIM 70093T displays the highest resistance to salt (up to 25%) GSK-3 described for Corynebacterium so far. Here we present a summary classification and a set of features for C. halotolerans YIM 70093T, together with the description of the genomic sequencing and annotation. Classification and features A representative genomic 16S rRNA sequence of C. halotolerans YIM 70093T was compared to the Ribosomal Database Project database [7], confirming the initial taxonomic classification.

cyanea, on the one hand, and S. viridis, S. azurea and S. marina, on the other hand, was generated with the Genome-to-Genome Distance Calculator (GGDC) [56-58]. This system calculates the distances by comparing the genomes to obtain HSPs (high-scoring segment pairs) and interfering distances necessary via a set of formulas (1, HSP length / total length; 2, identities / HSP length; 3, identities / total length). For convenience the GGDC also reports model-based DDH estimates along with their confidence intervals [58]. Table 5 shows the results of the pairwise comparison. Table 5 Pairwise comparison of S. cyanea with S. viridis, S. azurea and S. marina using the GGDC (Genome-to-Genome Distance Calculator). The comparison of S. cyanea with S. azurea reached the highest scores using the GGDC, 71% of the average of genome length are covered with HSPs.

The identity within the HSPs was 85%, whereas the identity over the whole genome was 61%. The lowest similarity scores were observed in the comparison of S. cyanea with S. marina with only 28% of the average of both genome lengths covered with HSPs. The identity within these HSPs was 79%, whereas the identity over the whole genome was only 22%. With regard to S. cyanea and S. azurea the corresponding DDH estimates were below the 70% threshold under formulas 1-3 throughout: 52.6% (��3), 28.6% (��3) and 45.4% (��3). The DDH estimates confidence intervals are given in parentheses as provided by [58]. The remaining pairings resulted in even smaller DDH estimates (data not shown).

As expected, those distances relating HSP coverage (formula 1) and number of identical base pairs within HSPs to total genome length (formula 3) are higher between S. cyanea and S. azurea than between S. cyanea and S. viridis or S. marina, respectively. That the distances relating the number of identical base pairs to total HSP length (formula 2) behave differently indicates that the genomic similarities between all four type strain genomes are strongly restricted to more conserved sequences, a kind of saturation phenomenon [56]. In order to further compare the genomes of S. cyanea, S. viridis, S. azurea and S. marina, correlation values (Pearson coefficient) according to the similarity on the level of COG category, pfam and TIGRfam were calculated (see Table 6). The highest correlation value (0.97) was reached for S. cyanea and S.

azurea on the level of pfam data; the correlation values on the basis of COG and TIGRfam data were only slightly smaller with 0.96 and 0.93, respectively. As a correlation value of 1 indicates the highest correlation, we can find a very high correlation between the genomes of S. cyanea and S. azurea considering the above data [55]. Table 6 Carfilzomib Pearson’s correlation coefficients according to the similarity on the level of Pfam, COG category and TIGRfam (in this order and separated by slashes). The synteny dot plots in Figure 4 shows nucleotide-based comparisons of the genomes of S.

After the initial examination, a personal kit containing a new toothbrush (Leader?, Facilit Odontol��gica e Perfumaria Ltda., Rio de Janeiro, RJ, Brazil), and a test KRX-0401 or control gel was given to all participants. They were then instructed to apply the gel over all bristles of the toothbrush and brush their teeth for one minute, three times a day, using their habitual technique. Verbal and written instructions about the correct use of oral hygiene products were given to all subjects as well. In addition to verbal instructions, participants were given recommendations to follow at home. On the last day of the experiment stage (day 90), indices were recorded and the teeth were polished with pumice. Statistical analysis ANOVA and Student Newman-Keuls post-hoc analysis were performed to evaluate statistical differences between the control and test groups on days 0 and 90 (��=.

05). In each group, the mean scores of all indices were compared between baseline and the end of the trial with the paired t-test (��=.05). However, for illustrative purposes, the results are presented as means and standard deviations. RESULTS All participants completed the trial. Both test gels had good acceptance and did not produce any adverse effects, such as ulcerations or allergic reactions. At baseline, there was no statistically significant difference between the control and test groups with respect to mean PI (P=.8376) and BI (P=.3198). These findings indicated that all groups were well balanced at baseline (Tables 1 and and2).2). At day 90, there was a statistically significant difference in PI and BI scores between the control and test groups (P<.

05) (Table 1). Table 1 Plaque index (PI) on day 0 and day 90, control and test groups (mean + standard deviation). Table 2 Bleeding index (BI) on day 0 and day 90, control and test groups (mean �� standard deviation). Comparison of means between baseline and day 90 in each group showed a statistically significant difference in BI and PI scores for the CLX and LS groups in relation to the control group (P<.05), but no difference between the CLX and LS groups (P>.05) (Tables 1 and and22). DISCUSSION The inability of the adult population to perform adequate mechanical tooth cleaning has stimulated the search for chemotherapeutic agents that can improve plaque control and gingivitis.

4,5,6,7 This paper presents the data of a clinical study where a phytopharmaceutical agent in gel dentifrice formulation was used in a group of patients with gingivitis and compared GSK-3 with chlorhexidine digluconate. The design was based on previous studies and it was chosen to generate the best possible clinical evidence.14,25 Chlorhexidine digluconate has been tested for many years and its long-term efficacy and safety have been confirmed in several in vivo studies.

1 unit. The highest pH at which cell-cell mostly membrane fusion was induced in cells infected with wild-type virus was 5.9 (Fig. (Fig.2).2). The H241Q and K582I mutations reduced the pH of membrane fusion to 5.6 and 5.4, respectively, while the Y231H and N1142K mutations increased the pH of membrane fusion to 6.3 and 6.4, respectively. The N1142K mutation, which increased the pH of HA activation by 0.5 pH unit, resulted in decreased virus fitness over several cycles of replication in vitro, while the other mutations did not alter in vitro replication kinetics. FIG. 2. The pH of HA-mediated membrane fusion by wild-type and mutant H5N1 influenza viruses in Vero cells was measured at 0.1 pH increments and is expressed as the highest pH value at which syncytium formation was observed.

The N1142K mutation is genetically unstable over multiple passages in eggs. To test the genetic stability of the HA protein mutations, wild-type and mutant viruses were passaged 10 times in 10-day-old embryonated chicken eggs. The sequence identity of each of the passage 1 (P1) recombinant viruses had been confirmed previously (36). Purified viral RNA sampled from allantoic fluid at P5 and P10 was sequenced. In parallel, syncytium formation assays were performed using Vero cells infected with P10 viruses to determine whether repeated passage in eggs resulted in any mutations that might alter the acid stability of the viral HA proteins (Table (Table1).1). Wild-type virus and viruses containing the HA protein mutation H241Q or K582I showed no additional mutations over the course of 10 passages and no change in the pH of membrane fusion.

The virus containing the Y231H mutation maintained the mutation for at least 5 passages in eggs and acquired an additional HA protein mutation, R2281I, between P5 and P10. Residue R228 (H5 numbering) is located in the receptor-binding pocket of the HA1 subunit with its side chain facing away from the pocket (46, 57) such that the R2281I mutation may enhance receptor binding in eggs (56). Despite the extra R2281I mutation, the P10 virus caused membrane fusion at a pH of 6.3, as did P1 Y231H virus without the additional R2281I mutation. The virus containing the N1142K mutation in the HA protein was the only recombinant virus whose pH of membrane fusion was altered at P10 from that for the P1 stock virus, a decrease from pH 6.4 to 6.1 (Table (Table1).

1). Both P5 and P10 K1142N viruses showed reversion mutations, demonstrating that the N1142K mutation was not genetically stable and was selected against within 5 passages. TABLE 1. Genetic stability of recombinant H5N1 influenza viruses containing HA protein mutations after serial passages Dacomitinib in embryonated chicken eggs Changes in the pH of activation of the HA protein can alter the environmental stability of H5N1 influenza viruses.

Using an alternative higher specificity rule (2/3 analysis), according to the manufacturer��s suggestion, we observed a specificity of 98.9%. Only 1% (1/92) of the NED group was SEPT9 positive, so this method is more suitable to detect the NED samples correctly as healthy compared with 1/3 analysis. Applying this rule to the CRC cases, 73 samples or 79.3% showed SEPT9 positivity, a level of sensitivity http://www.selleckchem.com/products/XL184.html that outperforms gFOBT and CEA for the detection of CRC. The selection of a higher sensitivity or higher specificity algorithm may be driven by differing objectives of screening programs. In this study, we also compared the sensitivity of methylated Septin 9 as a serum biomarker for both left- and right-sided CRC to gFOBT and CEA serum level.

Recent studies have demonstrated differences in the molecular patterns of colorectal carcinogenesis based on factors such as age, gender, and tumor localization [27]�C[30]. The elevated number of diagnosed left-sided colon cancers may be due to anatomical factors such as the lower colonic lumen diameter. Ghazi et al. found that left-sided CRC tended to have a lower T stage and higher N stage. However, they found no significant difference in the number of involved lymph nodes between the colonic locations. In addition, right-sided CRC has a worse prognosis than left-sided [31] and Weiss et al. found a higher mortality rate in right-sided stage III CRC [32]. While right-sided tumors display elevated gene expression levels of cell cycle control and Wnt signaling genes, left-sided colon cancers show reduced expression of tumor suppressor genes and cytokeratin 20 and elevated expression of COX-1 and genes that promote stromal expansion [33], [34].

Furthermore, difference in tumorigenesis between left- and right-sided colon cancers may be caused by epigenetic factors, as shown by differences in methylation. In regard to DNA methylation, it is known that some right-sided (CIMP) colon cancers have more frequent alterations as compared to left-sided cancers. Left-sided colon cancers can display a mutator phenotype, while right-sided tumors display as hypermethylator phenotype [35]�C[37]. Taken together, it is of significant clinical importance that screening methods for CRC are reliable for both left- and right-sided CRC. CRC detection by gFOBT in the context of left and right side of the colon has been evaluated in previous studies.

Steele et al. found gFOBT to be less sensitive for Carfilzomib both rectal cancer and right-sided cancer of the colon [38]. In our study, more left-sided CRC (83%) cases were detected by gFOBT than right-sided CRCs (50%) as well. The reason for detecting more left sided CRC may be explained by the localization of the disease and, due to stool consistency, blood originating from left-sided cancers may appear earlier in the feces.

81 (57%) of the cases and120 samples were wild-type (43%). We then we performed a second independent mutation analysis on all samples selleck products and observed that the results of the BRAF/KRAS and PIK3CA/NRAS assays were completely reproducible. We found 130 KRAS, 32 PIK3CA, 13 BRAF and 6 NRAS mutations. Details of the mutations are given in table 1. Of the 32 PIK3CA mutations 12 were single mutations, 19 occurred together with a KRAS mutation and 1 with a BRAF mutation. The mutation assays were performed in a blinded fashion and the results were compared afterwards with the results of the sequence analysis. There were 14 samples with discrepant results between sequence analysis for KRAS exon 2 (covering codons 12 and 13) and the BRAF/KRAS mutation assay.

As the latter had already been confirmed by an independent assay, the 14 samples were resequenced. In 9 cases, the sequence outcome now appeared identical to the mutation assay result and in 4 cases sequencing was unsuccessful due to poor DNA quality. In the 1 remaining sample (no. 289) sequencing suggested wild-type whereas the mutation assay detected a G12V mutation with the mutant peak being 6% of the wild type peak, suggesting that the tumor was heterogeneous. Figure 3 depicts a concise overview of sequence and mutation assay results. A detailed overview of the results of sequencing and mutation assays is given in Supplementary Table S3. In this table we also included the relative peak height of the mutant peaks compared to the wild type peak as observed in the mutation assays.

Note that this is at best semi-quantitative because of the different absorbances of the fluorescent labels, however, it gives an indication of the relative proportions of mutant and wild-type genes. Wild-type peaks in the assays have a height between 2000�C8000. Peaks (mutant) with a peak height of 100 are always visible. Based on this we estimate that sensitivity is between 1�C5%. This correlates well with the sensitivity calculated from dilution experiments in a similar assay as published previously [13]. Note that in some samples the mutant KRAS peaks were much higher than the peaks representing the wild type allele. We presume that this indicates loss of the wild-type allele. Figure 3 Overview of the resultsobtained by the mutation assays (A) and by sequence analysis (B) in 294 tumor DNA samples from patients with advanced colorectal cancer.

GSK-3 Table 1 Mutations identified with the BRAF/KRAS and PIK3CA/NRAS assays. Costs Comparison Next we compared the costs of both approaches. We observed mutations in all 4 genes, hence we assume that future mutation analysis, be it by sequencing or any other technique, will include the PIK3CA, NRAS and BRAF genes. We calculated the costs for all materials and reagents including those associated with the running of samples on the ABI 3130 XL sequencer. These amounted to � 7.03 for the two mutation assays together and � 59.54 for bi-directional sequencing of the 7 exons. Details are given in Sup