Custom Genomic solutions in India - Biotechdesk

What is it?

Asymmetrical PCR amplification can selectively or randomly amplify any unknown DNA sequence. This is accomplished in two quick steps:

1. An asymmetrical linear PCR step using Degenerate Random Tag (DRT) DNA primers followed by
2. A regular PCR step using a specific and a tagging prime.
   
   

It is extremely quick, inexpensive, reliable and simple, making it the ideal solution for genomic DNA amplification and cloning.

Transgenics

APA facilitates the tracking of transgene(s) in non-transient cell lines. A crucial element of this technology is that it can reveal the location and orientation of the transgene in correlation to a selected marker gene, and the accurate copy number.

Diagnostics

The most promising application of APA Technology is the newly emerging field of molecular diagnostics based on biochip and DNA microarray technology. The opportunity of amplifying DNA from multiple pathogens and screening them in parallel using APA Technology in conjunction with DNA microarray, will find increasing acceptance in the food, environmental, clinical and blood bank screening sectors.

BAC sequencing

BAC clone end sequencing provides valuable clues for contig alignment and generates anchor sequences with multiple starting points-enabling sequence walking. Usually, BAC end sequencing is limited by two factors: 1) a large amount of DNA is required, 2) the BAC vector is typically a single copy vector, making it extremely hard to extract large amounts of DNA from a BAC clone. With our innovative APA approach, only 2-5 ng of template DNA are needed for each reaction, almost 1000 times less than required by the conventional procedure! Also, APA significantly enhances the quality of sequencing data. A conclusive proof for this claim is the ability to sequence up to 1000 nucleotides in a single reaction.

Sequencing strategies

Our powerful APA technology platform provides a variety of innovative solutions to genome sequencing, as well as superior performance and reliability when compared with the archaic and ineffective shotgun sequencing methodology. Using our full genome sequencing strategy, all the BAC clones are sequenced from both ends.


The end sequencing data will be used for BAC clone contig alignment and the relevant end sequences will also serve as anchor sequences for bi-directional genome walking. This concurrent approach facilitates proof reading of the newly sequenced data. These properties translate into a validated complete genome sequence without gaps.

Gap Filling

One of the most effective uses of APA Technology is Gap filling in genome integration. Conventional Gap filling is often laborious, time consuming and ineffective. Through APA Technology, the sequencing process to walk the BAC clones is extremely simple and reliable with the additional advantage that only minute amount of DNA is required. In cases where conventional sequencing using BAC clones is not possible, APA Technology offers the unique advantage of sequencing directly from the genomic DNA.

Transcriptomics

APA Technology's best application to date is that it enables scientists in the generation of full-length cDNA transcripts and facilitates 5' and 3' RACE. This confers a unique advantage in understanding gene regulatory pathways. APA Technology is thus a crucial tool in delineating gene function and regulation domains in the Human Transcriptome.

Please also take a look at Custom genomic solutions and BAC library construction services from our company.

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