NDLI: Updated instrumentation for continuous array genotyping of short insertion/deletion polymorphisms

Content Provider	IEEE Xplore Digital Library
Author	Rusch, T.L. Dickinson, W. Che, J. Fieweger, K. Chudyk, J. Doktycz, M.J. Yu, A. Weber, J.L.
Copyright Year	2003
Description	Author affiliation: Marshfield Clinic Res. Foundation, WI, USA (Rusch, T.L.; Dickinson, W.; Che, J.; Fieweger, K.; Chudyk, J.)
Abstract	Methods for DNA testing are abundant, with much emphasis placed on short tandem repeat polymorphisms (STRPs) and single nucleotide polymorphisms (SNPs). However, lower cost, higher-throughput genotyping is still needed. Another important polymorphism is the diallelic short insertion/deletion (indel). Our laboratory is using a continuous reel of 384 well arrays on polypropylene tape to genotype indel polymorphisms. The reel of continuous arrays allows lower cost automation, and decreased reaction volumes. The diallelic indel is typed by tagging allele-specific PCR primers with a FAM or JOE molecular beacon uniprimer. Our most recent tape has wells with reaction volumes of 700 nanoliters or less. Commercial equipment for the microtape is not available. A series of instruments to handle the tape was developed in-house. A pipetting instrument was developed to deliver the specific DNA samples or other reagents. A solenoid microvalve aspirating and jetting unit was developed for dispensing a common reaction mix. The arrays are sealed with commercially available continuous roll seal material prior to polymerase chain reaction (PCR). After PCR the arrays are scanned using an epi-fluorescence detection unit. The detector uses an argon ion laser for excitation and photomultiplier tubes (PMTs) for detection.
Sponsorship	Whitaker Found
Starting Page	3582
Ending Page	3585
File Size	364420
Page Count	4
File Format	PDF
ISBN	0780377893
ISSN	1094687X
DOI	10.1109/IEMBS.2003.1280927
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2003-09-17
Publisher Place	Mexico
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	Instruments DNA Costs Testing Laboratories Automation Tagging Solenoids Microvalves Seals
Content Type	Text
Resource Type	Article

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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