Ultra-rapid DNA analysis using HyBeacon™ probes and direct PCR amplification from saliva

2002 ◽  
Vol 16 (5) ◽  
pp. 319-326 ◽  
Author(s):  
D.J. French ◽  
C.L. Archard ◽  
M.T. Andersen ◽  
D.G. McDowell
Keyword(s):  
Dna Analysis ◽  
Pcr Amplification ◽  
Direct Pcr ◽  
Rapid Dna Analysis

DNA analysis of cystic fibrosis in Brazil by direct PCR amplification from Guthrie cards

1993 ◽  
Vol 46 (6) ◽  
pp. 665-669 ◽  
Author(s):  
S. Raskin ◽  
J. A. Phillips ◽  
M. R. S. Krishnamani ◽  
C. Vnencak-Jones ◽  
R. A. Parker ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Dna Analysis ◽  
Pcr Amplification ◽  
Direct Pcr ◽  
Guthrie Cards

scholarly journals Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs

2012 ◽  
Vol 64 (3) ◽  
pp. 851-858 ◽  
Author(s):  
Natasa Kovacevic-Grujicic ◽  
S. Davidovic ◽  
Dijana Takic ◽  
Marija Mojsin ◽  
Milena Stevanovic
Keyword(s):  
Mitochondrial Dna ◽  
Dna Analysis ◽  
Pcr Amplification ◽  
Buccal Cell ◽  
Buccal Cells ◽  
Direct Pcr ◽  
Long Term Storage ◽  
Short Period ◽  
Term Storage

Amplification of human mitochondrial DNA (mtDNA) has been widely used in population genetics, human evolutionary and molecular anthropology studies. mtDNA hypervariable segments I and II (HVSI and HVSII) were shown to be a suitable tool in genetic analyses due to the unique properties of mtDNA, such as the lack of recombination, maternal mode of inheritance, rapid evolutionary rate and high population-specific polymorphisms. Here we present a rapid and low-cost method for direct PCR amplification of a 330 bp fragment of HVSI from buccal cell samples. Avoiding the DNA isolation step makes this method appropriate for the analysis of a large number of samples in a short period of time. Since the transportation of samples and fieldwork conditions can affect the quality of samples and subsequent DNA analysis, we tested the effects of long-term storage of buccal cell swabs on the suitability of such samples for direct PCR amplification. We efficiently amplified a 330 bp fragment of HVSI even after the long-term storage of buccal cells at room temperature, +4?C or at -20?C, for up to eight months. All examined PCR products were successfully sequenced, regardless of sample storage time and conditions. Our results suggest that the direct PCR amplification of the HVSI region from buccal cells is a method well suited for large-scale mtDNA population studies.

scholarly journals DMSO Improves the Ski-Slope Effect in Direct PCR

2021 ◽  
Vol 11 (4) ◽  
pp. 1943
Author(s):  
Joo-Young Kim ◽  
Ju Yeon Jung ◽  
Da-Hye Kim ◽  
Seohyun Moon ◽  
Won-Hae Lee ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Analytical Techniques ◽  
Direct Pcr ◽  
Dna Profile ◽  
Novel Technologies ◽  
Specific Amplification ◽  
Polymerase Chain ◽  
Dna Profiles

Analytical techniques such as DNA profiling are widely used in various fields, including forensic science, and novel technologies such as direct polymerase chain reaction (PCR) amplification are continuously being developed in order to acquire DNA profiles efficiently. However, non-specific amplification may occur depending on the quality of the crime scene evidence and amplification methods employed. In particular, the ski-slope effect observed in direct PCR amplification has led to inaccurate interpretations of the DNA profile results. In this study, we aimed to reduce the ski-slope effect by using dimethyl sulfoxide (DMSO) in direct PCR. We confirmed that DMSO (3.75%, v/v) increased the amplification yield of large-sized DNA sequences more than that of small-sized ones. Using 50 Korean buccal samples, we further demonstrated that DMSO reduced the ski-slope effect in direct PCR. These results suggest that the experimental method developed in this study is suitable for direct PCR and may help to successfully obtain DNA profiles from various types of evidence at crime scenes.

scholarly journals Direct PCR amplification of DNA from human bloodstains, saliva, and touch samples collected with microFLOQ ® swabs

2018 ◽  
Vol 32 ◽  
pp. 80-87 ◽  
Author(s):  
Angie Ambers ◽  
Rachel Wiley ◽  
Nicole Novroski ◽  
Bruce Budowle
Keyword(s):  
Pcr Amplification ◽  
Direct Pcr

A modified direct PCR amplification method using the GlobalFiler™ PCR Amplification Kit on bloodstains collected using microFLOQ™ direct swabs

2019 ◽  
Vol 7 (1) ◽  
pp. 30-32
Author(s):  
Yongxun Wong ◽  
Boon Kiat Ng ◽  
Kevin Wai Yin Chong ◽  
Wei Siong Holden Lim ◽  
Afiqah Razanah Rosli ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Direct Pcr ◽  
Amplification Method

scholarly journals Numerous uncharacterized and highly divergent microbes which colonize humans are revealed by circulating cell-free DNA

2017 ◽  
Vol 114 (36) ◽  
pp. 9623-9628 ◽  
Author(s):  
Mark Kowarsky ◽  
Joan Camunas-Soler ◽  
Michael Kertesz ◽  
Iwijn De Vlaminck ◽  
Winston Koh ◽  
...  
Keyword(s):  
Human Body ◽  
Sequence Data ◽  
Human Microbiome ◽  
Pcr Amplification ◽  
The Body ◽  
Direct Pcr ◽  
Cell Free Dna ◽  
Free Dna ◽  
Novel Taxa ◽  
Circulating Cell Free Dna

Blood circulates throughout the human body and contains molecules drawn from virtually every tissue, including the microbes and viruses which colonize the body. Through massive shotgun sequencing of circulating cell-free DNA from the blood, we identified hundreds of new bacteria and viruses which represent previously unidentified members of the human microbiome. Analyzing cumulative sequence data from 1,351 blood samples collected from 188 patients enabled us to assemble 7,190 contiguous regions (contigs) larger than 1 kbp, of which 3,761 are novel with little or no sequence homology in any existing databases. The vast majority of these novel contigs possess coding sequences, and we have validated their existence both by finding their presence in independent experiments and by performing direct PCR amplification. When their nearest neighbors are located in the tree of life, many of the organisms represent entirely novel taxa, showing that microbial diversity within the human body is substantially broader than previously appreciated.

scholarly journals Direct PCR amplification of HCV RNA from human serum.

1992 ◽  
Vol 1 (4) ◽  
pp. 291-292 ◽  
Author(s):  
A Ravaggi ◽  
D Primi ◽  
E Cariani
Keyword(s):  
Human Serum ◽  
Pcr Amplification ◽  
Hcv Rna ◽  
Direct Pcr

scholarly journals Rapid Detection of Phytophthora infestans in Late Blight-Infected Potato and Tomato Using PCR

Plant Disease ◽  
1997 ◽  
Vol 81 (9) ◽  
pp. 1042-1048 ◽  
Author(s):  
C. L. Trout ◽  
J. B. Ristaino ◽  
M. Madritch ◽  
T. Wangsomboondee
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Pcr Amplification ◽  
Accurate Method ◽  
Phytophthora Species ◽  
Universal Primers ◽  
Direct Pcr ◽  
Field Samples ◽  
Pcr Products ◽  
Oomycete Pathogen

Late blight caused by the oomycete pathogen Phytophthora infestans is a devastating disease of potato and tomato worldwide. A rapid and accurate method for specific detection of P. infestans is necessary for determination of late blight in infected fruit, leaves, and tubers. Ribosomal DNA (rDNA) from four isolates of P. infestans representing the four genotypes US1, US6, US7, and US8 was amplified using polymerase chain reaction (PCR) and the universal primers internal transcribed spacer (ITS) 4 and ITS5. PCR products were sequenced using an automated sequencer. Sequences were aligned with published sequences from 5 other Phytophthora species, and a region specific to P. infestans was used to construct a PCR primer (PINF). Over 140 isolates representing 14 species of Phytophthora and at least 13 other genera of fungi and bacteria were used to screen the PINF primer. PCR amplification with primers PINF and ITS5 results in amplification of an approximately 600 base pair product with only isolates of P. infestans from potato and tomato, as well as isolates of P. mirabilis and P. cactorum. P. mirabilis and P. cactorum are not pathogens of potato; however, P. cactorum is a pathogen of tomato. P. infestans and P. cactorum were differentiated by restriction digests of the amplified product. The PINF primer was used with a rapid NaOH lysis technique for direct PCR of P. infestans from infected tomato and potato field samples. The PINF primer will provide a valuable tool for detection of P. infestans in potatoes and tomatoes.

Optimization of multiplexed PCR on an integrated microfluidic forensic platform for rapid DNA analysis

The Analyst ◽  
2012 ◽  
Vol 137 (23) ◽  
pp. 5510 ◽  
Author(s):  
Matthew D. Estes ◽  
Jianing Yang ◽  
Brett Duane ◽  
Stan Smith ◽  
Carla Brooks ◽  
...  
Keyword(s):  
Dna Analysis ◽  
Multiplexed Pcr ◽  
Rapid Dna Analysis
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