Phylogenetic analysis shows that New Zealand isolates of Neonectria ditissima are similar to European isolates

Neonectria ditissima causes a debilitating apple tree canker disease. We determined the efficacy of polymerase chain reaction primers, originally designed for European strains, by sequencing New Zealand strains. The concatenated ribosomal inter-transcribed spacer and β-tubulin gene regions of 17 New Zealand isolates were compared with those of two European strains by phylogenetic analysis. New Zealand and European isolates of N. ditissima were in the same clade, suggesting that there has been little change in these gene regions following introduction to New Zealand. There was 100% homology with Bt-FW135 and Bt-RW284 primers. Based on sequencing 17 New Zealand isolates from several locations, these polymerase chain reaction primers can be relied upon to amplify New Zealand isolates of N. ditissima.

Myriad post-Myriad

The US Supreme Court’s 2013 decision, holding patent claims to isolated, endogenous deoxyribonucleic acid (DNA) sequences to be invalid, seemed to have limited negative impact on Myriad Genetics whose patent on the isolated BRCA1 and BRCA2 genes were at the heart of the case. This article explains this minimal impact in two ways. First, the Court’s decision still left synthetic DNA patentable, leaving that as a fruitful source for commercialization by companies like Myriad. The Federal Circuit’s subsequent decision, however, invalidated Myriad’s product claims over the synthetic polymerase chain reaction primers based on the isolated DNA sequences. Second, the Court’s decision did not address the patentability of mined genetic data for diagnostic and therapeutic purposes. This field of genetic data mining is precisely where Myriad has moved in its patenting activity.

High-throughput retrotransposon-based genetic diversity of maize germplasm assessment and analysis

Maize is one of the world’s most important crops and a model for grass genome research. Long terminal repeat (LTR) retrotransposons comprise most of the maize genome; their ability to produce new copies makes them efficient high-throughput genetic markers. Inter-retrotransposon-amplified polymorphisms (IRAPs) were used to study the genetic diversity of maize germplasm. Five LTR retrotransposons (Huck, Tekay, Opie, Ji, and Grande) were chosen, based on their large number of copies in the maize genome, whereas polymerase chain reaction primers were designed based on consensus LTR sequences. The LTR primers showed high quality and reproducible DNA fingerprints, with a total of 677 bands including 392 polymorphic bands showing 58% polymorphism between maize hybrid lines. These markers were used to identify genetic similarities among all lines of maize. Analysis of genetic similarity was carried out based on polymorphic amplicon profiles and genetic similarity phylogeny analysis. This diversity was expected to display ecogeographical patterns of variation and local adaptation. The clustering method showed that the varieties were grouped into three clusters differing in ecogeographical origin. Each of these clusters comprised divergent hybrids with convergent characters. The clusters reflected the differences among maize hybrids and were in accordance with their pedigree. The IRAP technique is an efficient high-throughput genetic marker-generating method.

A set of experimentally validated, mutually orthogonal primers for combinatorially specifying genetic components

The design and synthesis of novel genes and deoxyribonucleic acid (DNA) sequences is a central technique in synthetic biology. Current methods of high throughput gene synthesis use pooled oligonucleotides obtained from custom-designed DNA microarray chips, and rely on orthogonal (non-interacting) polymerase chain reaction primers to specifically de-multiplex, by amplification, the precise subset of oligonucleotides necessary to assemble a full length gene. The availability of a large validated set of mutually orthogonal primers is therefore a crucial reagent for high-throughput gene synthesis. Here, we present a set of 166 20-nucleotide primers that are experimentally verified to be non-interacting, capable of specifying 13 695 unique genes. These primers represent a valuable resource to the synthetic biology community for specifying genetic components that can be assembled through a scalable and modular architecture.