On-Chip Isothermal Polymerase Chain Reaction

2007 ◽  
Author(s):  
Alexandre Persat ◽  
Tomoyuki Morita ◽  
Juan G. Santiago
Keyword(s):  
Concentration Field ◽  
Chain Reaction ◽  
Base Pairs ◽  
Denaturant Concentration ◽  
Control Scheme ◽  
Polymerase Chain ◽  
Spatio Temporal ◽  
On Chip ◽  
Spatially Varying ◽  
Dna Template

We present a novel technique for on-chip PCR where temperature is held constant and uniform in the reactor. Specific chemicals, known as denaturants, have the ability to melt DNA. A flow control scheme establishes spatio-temporal fluctuations in the concentration of denaturants along a microchannel, while electromigration drives DNA through this spatially varying denaturant concentration field. Preliminary results show denaturation and extension of a 200 base pairs (bp) DNA template.

Detection of the Glanzmann's Thrombasthenia Mutations in Arab and Iraqi-Jewish Patients by Polymerase Chain Reaction and Restriction Analysis of Blood or Urine Samples

1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman
Keyword(s):  
Polymerase Chain Reaction ◽  
Base Pair ◽  
Restriction Analysis ◽  
Urine Samples ◽  
Chain Reaction ◽  
Base Pairs ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Base Pair Deletion ◽  
Polymerase Chain

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.

On-chip polymerase chain reaction microdevice employing a magnetic droplet-manipulation system

2008 ◽  
Vol 130 (2) ◽  
pp. 583-588 ◽  
Author(s):  
Hiroyoshi Tsuchiya ◽  
Mina Okochi ◽  
Nobuhiro Nagao ◽  
Mitsuhiro Shikida ◽  
Hiroyuki Honda
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Manipulation System ◽  
Polymerase Chain ◽  
Droplet Manipulation ◽  
On Chip ◽  
Magnetic Droplet

Molecular cloning, characterization, and expression analysis of a novel sema-2a homologue in Polyrhachis vicina (Hymenoptera: Formicidae)

2010 ◽  
Vol 142 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Jing Luo ◽  
Geng-Si Xi ◽  
Shu-Min Lü ◽  
Ke Li ◽  
Qing Li
Keyword(s):  
Untranslated Region ◽  
Axonal Guidance ◽  
Mrna Levels ◽  
Chain Reaction ◽  
Base Pairs ◽  
Reading Frame ◽  
Acid Protein ◽  
Polymerase Chain ◽  
Polyrhachis Vicina ◽  
Amino Acid Protein

AbstractThe semaphorin gene family plays important roles in axonal guidance in vertebrates and invertebrates. Semaphorin 2a, a member of the semaphorin family, belongs to class 2, which is found only in invertebrates. In our study, semaphorin 2a was cloned from the ant Polyrhachis vicina Roger. The full length of P. vicina semaphorin 2a (Pv-sema-2a) is 2763 base pairs (bp) and it contains a 5′-untranslated region (UTR) 92 bp long and a 3′-UTR 521 bp long. The open reading frame of Pv-sema-2a encodes a 716-amino-acid protein with a predicted molecular mass of 81.1 kilodaltons. Real-time quantitative reverse-transcription – polymerase chain reaction indicated that Pv-sema-2a mRNA is differentially expressed during P. vicina development, in the whole bodies as well as the heads of different castes. The high mRNA levels in embryos and pupae suggest that Pv-sema-2a plays an important role in ant development.

VARIAN NON-DELESI 9 PASANG BASA DNA MITOKONDRIA MANUSIA SAMPEL FORENSIK BALI

2005 ◽  
Vol 6 (1) ◽  
pp. 74
Author(s):  
Gun Gun Gumilar ◽  
A. Saifuddin Noer
Keyword(s):  
Gel Electrophoresis ◽  
Secondary Data ◽  
Primary Data ◽  
Agarose Gel Electrophoresis ◽  
Chain Reaction ◽  
Base Pairs ◽  
Human Mitochondrial Dna ◽  
Deletion Variant ◽  
Forensic Samples ◽  
Polymerase Chain

One of human mitochondrial DNA (mtDNA) variant is a 9 base pairs (bp) deletion in the COII/tRNALys intergenic region. In construction mtDNA nomenclature, 9-bp deletion database consist of primary and secondary data is needed, including Bali bombing forensic samples. Here we report a 9-bp non- deletion mtDNA variant from Bali bombing forensic samples to complete primary data. Polymerase Chain Reaction (PCR) technique with 2 set primer was used to detect 9-bp deletion. The PCR result was detected by agarose gel electrophoresis, which showed two bands (0.1 and 0.4 kb) for non-deletion variant control, and one band (0.4 kb) for deletion variant control. If the sample has 9-bp deletion, only one of the primer pairs could amplify a fragment of 0.4 kb. If the sample does not have 9-bp deletion, the other primer pair will amplify a 0.1 kb product. The result showed that none of the 24 samples has 9-bp deletion. These results are contributed to the human mtDNA database and nomenclature construction. Keywords: mtDNA, 9-bp deletion, PCR

A Polymerase Chain Reaction (PCR) Method for Sex and Species Determination with Novel Controls for Deoxyribonucleic Acid (DNA) Template Length

1992 ◽  
Vol 37 (1) ◽  
pp. 13207J ◽  
Author(s):  
R. E. Gaensslen ◽  
Karen M. Berka ◽  
Dina A. Grosso ◽  
Gualberto Ruano ◽  
Elaine M. Pagliaro ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Deoxyribonucleic Acid ◽  
Chain Reaction ◽  
Species Determination ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Dna Template

Specific detection of mousepox virus by polymerase chain reaction

1997 ◽  
Vol 31 (3) ◽  
pp. 201-205 ◽  
Author(s):  
Heinrich Neubauer ◽  
Martin Pfeffer ◽  
Hermann Meyer
Keyword(s):  
Polymerase Chain Reaction ◽  
Rapid Identification ◽  
Mouse Lung ◽  
Chain Reaction ◽  
Base Pairs ◽  
Viral Origin ◽  
Body Protein ◽  
Polymerase Chain ◽  
Mouse Lung Tissue ◽  
Type Inclusion

Polymerase chain reaction was applied to the rapid identification and detection of mousepox virus. This was accomplished by selection of primers targeting the A-type inclusion body protein gene. By investigating 20 strains belonging to five different species of the genus Orthopoxvirus, amplification was achieved only with the seven mousepox virus strains examined. The size of the resulting DNA fragment accounted for 116 base pairs and contained a recognition site for the restriction enzyme HindII, thus confirming its viral origin. Amplification of mousepox virus specific sequences was also possible from infected mouse lung tissue and serum.

Portable plastic syringe as a self-actuated pump for long-distance uniform delivery of liquid inside a microchannel and its application for flow-through polymerase chain reaction on chip

RSC Advances ◽  
2015 ◽  
Vol 5 (16) ◽  
pp. 12071-12077 ◽  
Author(s):  
Wenming Wu ◽  
Kieu The Loan Trinh ◽  
Yu Zhang ◽  
Nae Yoon Lee
Keyword(s):  
Polymerase Chain Reaction ◽  
Flow Rate ◽  
Chain Reaction ◽  
Long Distance ◽  
Chip Flow ◽  
Hands On ◽  
Plastic Syringe ◽  
Flow Through ◽  
Polymerase Chain ◽  
On Chip

A strategy for realizing self-actuated pumping with uniform flow rate over a long distance is introduced using hands-on operation of disposable syringe, and was applied for on-chip flow-through PCR inside a serpentine PMMA microchannel.

scholarly journals Barcode DNA Edelweis (Anaphalis javanica) Berdasarkan Gen matK

Jurnal MIPA ◽  
2015 ◽  
Vol 4 (2) ◽  
pp. 131
Author(s):  
Muzakir Rahalus ◽  
Maureen Kumaunang ◽  
Audy Wuntu ◽  
Julius Pontoh
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Dna Barcode ◽  
Living Organism ◽  
Chain Reaction ◽  
Base Pairs ◽  
Barcode Dna ◽  
Reverse Primer ◽  
Total Dna ◽  
Polymerase Chain

DNA barcode merupakan metode identifikasi organisme hidup dengan menggunakan urutan DNA pendek (± 500 pasang basa). Tujuan dari penelitian ini adalah memperoleh barcode DNA Edelweis dan menganalisis kemiripan gen matK Edelweis (Anaphalis javanica) dengan kerabat terdekatnya. Isolasi DNA total Edelweis berhasil dilakukan dengan menggunakan  manual prosedur dari InnuPrep Plant DNA Kit yang dimodifikasi. Gen matK parsial telah diisolasi dengan metode Polymerase Chain Reaction (PCR) menggunakan Primer forward matK-1RKIM-f dan Primer Reverse matK-3FKIM-r. Hasil analisis sekuens menghasilkan barcode DNA edelweis berukuran 843 bp. Hasil analisis kemiripan menunjukkan tingkat kekerabatan terdekat dengan A. margaritaceae yaitu 99.86% pada BOLD System dan 100 % pada NCBI.DNA barcode is a method to identify living organism by using several short sequences of DNA (± 500 base pairs). The purpose of this study was to obtain a DNA barcode and analyze the similarity of matK genes of edelweis (Anaphalis javanica) with its closest relatives. Isolation of total DNA of edelweis has been succesfully done by using modified manual procedures of InnuPrep Plant Kit. matK partial gene has been isolated by the method of Polymerase Chain Reaction (PCR) using forward primer MATK-1RKIM-f and reverse primer MATK-3FKIM-r. Analysis of DNA sequences of edelweis confirmed its DNA barcode size was 843 bp. Furthermore, A. javanica showed similarity 99.86% in BOLD system and 100% in NCBI with A. margaritaceae.

scholarly journals Deconstructing the Polymerase Chain Reaction II: an improved workflow and effects on artifact formation and primer degeneracy

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7121
Author(s):  
Ankur Naqib ◽  
Silvana Poggi ◽  
Stefan J. Green
Keyword(s):  
Polymerase Chain Reaction ◽  
Community Structure ◽  
Microbial Community ◽  
Annealing Temperature ◽  
Degenerate Primer ◽  
Chain Reaction ◽  
Degenerate Primers ◽  
Polymerase Chain ◽  
Dna Template ◽  
Microbial Dna

Polymerase chain reaction (PCR) amplification of complex microbial genomic DNA templates with degenerate primers can lead to distortion of the underlying community structure due to inefficient primer-template interactions leading to bias. We previously described a method of deconstructed PCR (“PEX PCR”) to separate linear copying and exponential amplification stages of PCR to reduce PCR bias. In this manuscript, we describe an improved deconstructed PCR (“DePCR”) protocol separating linear and exponential stages of PCR and allowing higher throughput of sample processing. We demonstrate that the new protocol shares the same benefits of the original and show that the protocol dramatically and significantly decreases the formation of chimeric sequences during PCR. By employing PCR with annealing temperature gradients, we further show that there is a strong negative correlation between annealing temperature and the evenness of primer utilization in a complex pool of degenerate primers. Shifting primer utilization patterns mirrored shifts in observed microbial community structure in a complex microbial DNA template. We further employed the DePCR method to amplify the same microbial DNA template independently with each primer variant from a degenerate primer pool. The non-degenerate primers generated a broad range of observed microbial communities, but some were highly similar to communities observed with degenerate primer pools. The same experiment conducted with standard PCR led to consistently divergent observed microbial community structure. The DePCR method is simple to perform, is limited to PCR mixes and cleanup steps, and is recommended for reactions in which degenerate primer pools are used or when mismatches between primers and template are possible.

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