Abstract: Disclosed are methods for lysis of cells that combine three lysis steps—(1) heat, (2) detergent and (3) base—into a single step and that can be completed in a short period of time, e.g., a few minutes. The methods combine a normally incompatible detergent and base lysis, allow for simplified removal of detergent after lysis, and importantly, limits damage to DNA, such as shearing, that typically results from separate application of conventional lysis methods, yielding improved quality and quantities of genomic DNA (gDNA).

Abstract: Disclosed are methods for screening biological samples for the presence unknown microbes, such as bacteria and archaea or unknown eukaryotes using rRNA gene sequences or other highly conserved genetic regions, across multiple biological samples using a unique sequence tag (barcode) corresponding to the sample. The screening process tracks the unknown microbe or eukaryote in a diluted sample where the DNA has been prepared using whole genome amplification. The whole genome of the unknown microbe or eukaryote is then sequenced and assembled.

Abstract: Disclosed are methods for lysis of cells, such as bacteria present in microbiomes, that combine three lysis steps—(1) heat, (2) detergent and (3) base—into a single step and that can be completed in a short period of time, e.g., a few minutes. The methods combine a normally incompatible detergent and base lysis, allows for simplified removal of detergent after lysis, and importantly, yields improved quantities of genomic DNA (gDNA) from difficult to lyse bacteria.

Abstract: Disclosed are methods for screening biological samples for the presence unknown microbes, such as bacteria and archaea or unknown eukaryotes using rRNA gene sequences or other highly conserved genetic regions, across multiple biological samples using a unique sequence tag (barcode) corresponding to the sample. The screening process tracks the unknown microbe or eukaryote in a diluted sample where the DNA has been prepared using whole genome amplification. The whole genome of the unknown microbe or eukaryote is then sequenced and assembled.

Abstract: Disclosed are methods for lysis of cells, such as bacteria present in microbiomes, that combine three lysis steps—(1) heat, (2) detergent and (3) base—into a single step and that can be completed in a short period of time, e.g., a few minutes. The methods combine a normally incompatible detergent and base lysis, allows for simplified removal of detergent after lysis, and importantly, yields improved quantities of genomic DNA (gDNA) from difficult to lyse bacteria.

Abstract: Disclosed are compositions and methods for single-step PCR of a sample containing a complex target gene pool that can simultaneously amplify a wide variety of variant target gene sequences common to the sample while maintaining the original ratios of gene variants. The compositions and methods described herein utilize (1) a gene-specific primer pool that contains multiple variants that occur in a sample containing a complex mixture of target sequences that are both required for amplification of variants in the mixture which may introduce amplification bias, with (2) a non-specific PCR primer that is designed to target multiple gene-specific primer variants and eliminate amplification bias.

Abstract: Disclosed are methods and devices for lysing cells to release extract genomic DNA (gDNA). The methods use a mixture of microscopic glass beads and cells (for example, spores) that form a semi-dry cake that clings to a larger metal ball and the sides of the tube during bead beating lysis, greatly improving the efficiency of the bead beating process. The devices produce a chaotic motion which ensures that sufficient force is generated to open the cells, and that the metal ball impacts are distributed across the interior surface of the container so that all of the cell mixture is subjected to sufficient impacts to break the cells. As a result, spores and other difficult-to-lyse microbes, can be opened in seconds. The method reduces the number of steps and hands-on time by rapidly opening difficult to lyse cells, while preserving the integrity of the DNA.

Abstract: Disclosed are methods for lysis of cells, such as bacteria present in microbiomes, that combine three lysis steps—(1) heat, (2) detergent and (3) base—into a single step and that can be completed in a short period of time, e.g., a few minutes. The methods combine a normally incompatible detergent and base lysis, allows for simplified removal of detergent after lysis, and importantly, yields improved quantities of genomic DNA (gDNA) from difficult to lyse bacteria.

Abstract: Disclosed are methods for screening biological samples for the presence unknown microbes, such as bacteria and archaea or unknown eukaryotes using rRNA gene sequences or other highly conserved genetic regions, across multiple biological samples using a unique sequence tag (barcode) corresponding to the sample. The screening process tracks the unknown microbe or eukaryote in a diluted sample where the DNA has been prepared using whole genome amplification. The whole genome of the unknown microbe or eukaryote is then sequenced and assembled.

Abstract: Disclosed are methods for parallel single-step DNA purification starting with multiple crude biological samples for subsequent parallel PCR amplification of target DNA that attaches a unique DNA sequence tag (barcode) allowing all parallel processed samples to be combined into a single high-throughput sequencing run. The methods disclosed herein can be used to prepare and sequence dozens or hundreds of targeted samples as part of a rapid, highly parallel process, after which individual sample sequencing results are separated using the sample-specific tags (barcodes) to obtain results for each sample.

Abstract: A rapid method and kit is provided for identifying the targets of biologically active small molecules to identify new drugs that are capable of specific therapeutic effects as well as to identify novel small molecules including agonists and antagonists that may bind selected targets.

Abstract: Disclosed are methods for identifying nucleic acids in a sample of nucleic acids in which nucleic acids are initially unequal amounts. The methods include partitioning the starting population of nucleic acids to form one or more subpopulations, and then identifying nucleic acids that are present in different amounts in the partitioned nucleic acid sample as compared to the starting population. Also disclosed are methods of generating databases based on sizes of DNA fragments and size-based sequencing of those fragments.