A Method for Sequence-Independent in Vitro Amplification of DNA

University of Chicago
posted on 06/08/2009

• request more info 
•   
•  LICENSE ONLINE

Researchers at the University of Chicago have developed a method to amplify minute quantities of DNA in the sub pico gram (pg) range without prior knowledge of the sequence. Several micrograms of DNA can be obtained from as little as 0.1 pg of starting material.

• Amplification of microdissected chromosome fragments from any species for the generation of chromosome-specific or chromosome band-specific FISH probes and DNA libraries, which can then be used in downstream applications like high throughput sequencing approaches.
• Amplification of mini preparations of bacterial artificial chromosomes (BACs) or yeast artificial chromosomes (YACs) for the generation of FISH probes.
• Amplification of small traces of fragmented ancient DNA for high throughput sequencing.
• Amplification of small DNA traces in forensic settings for further analysis.
• Amplification of DNA from archival samples for further analysis.
• Amplification of DNA samples (eg. from patients or invironmental specimen) to increase the number of analyses that can be performed on each individual sample.
• Whole genome amplification of single cell genomes for pre-implantation genetic diagnostics: eg. the DNA of a single blastocyst cells or a single polar body cell can be amplified. Subsequently, extensive SNP typing to rule out aneuploidies for all chromosomes or testing for multiple genetic mutations could be performed without restrictions in DNA quantity. Currently this is not possible with any other technique.
• In combination with reverse transcription, to amplify the transcriptome of single cells with subsequent high throughput sequencing to determine single cell expression profiles.
• To monitor changes in the genome of single cells or small groups of cells (e.g. to monitor the mutational load in individual induced pluripotent stem (IPS) cell clones prior to expansion for therapeutic applications) using SIA followed by high throughput sequencing.
• Amplification of small DNA traces to identify unknown pathogens (eg. in the setting of suspected bioterrorism).
• Amplification of small amounts of DNA that contain secret messages or codes.

• The template DNA does not have to be purified.
• The method is extremely sensitive.
• It is a one-tube-procedure without the need for intermediate purification steps.
• There is no a priori sequence bias in the amplification reaction.