Direct amplification of casework samples in fabrics using GlobalFiler ® PCR Amplification Kit

2015 ◽  
Vol 5 ◽  
pp. e487-e489 ◽  
Author(s):  
P. Brito ◽  
N. Gouveia ◽  
V. Bogas ◽  
A. Serra ◽  
F. Balsa ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Direct Amplification

scholarly journals PCR Detection and RFLP Differentiation of Botrytis Species Associated with Neck Rot of Onion

Plant Disease ◽  
2002 ◽  
Vol 86 (6) ◽  
pp. 682-686 ◽  
Author(s):  
Karsten Nielsen ◽  
David S. Yohalem ◽  
Dan Funck Jensen
Keyword(s):  
Pcr Amplification ◽  
Pcr Detection ◽  
Sequence Characterized Amplified Region ◽  
Specific Pcr ◽  
Isolation Techniques ◽  
Detection And Identification ◽  
Fungal Dna ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Direct Amplification

Botrytis aclada and other Botrytis spp. can cause neck rot on onions, a storage disease that normally is very difficult to detect at harvest using traditional isolation techniques. Sequence characterized amplified region primers (BA2f/BA1r) were designed based on a previously cloned and amplified DNA fragment for direct amplification of isolates of Botrytis spp. associated with neck rot of onions. Digestion of the polymerase chain reaction (PCR) amplification product with the restriction enzyme ApoI makes it possible to distinguish the five groups: Botrytis aclada types AI and AII (B. allii); B. byssoidea; B. squamosa; and B. cinerea. The detection limit was 1 to 10 pg of pure fungal DNA. It was possible to detect B. aclada with the PCR method in artificially inoculated onion bulb tissue and in mature onion leaves showing no symptoms of the disease. The availability of a sensitive and specific PCR detection and identification method for Botrytis onion neck rot pathogens should facilitate ecological studies of this group of onion pathogens.

scholarly journals Analysis of Ammonia-Oxidizing Bacteria from Hypersaline Mono Lake, California, on the Basis of 16S rRNA Sequences

2000 ◽  
Vol 66 (7) ◽  
pp. 2873-2881 ◽  
Author(s):  
B. B. Ward ◽  
D. P. Martino ◽  
M. C. Diaz ◽  
S. B. Joye
Keyword(s):  
Denaturing Gradient Gel Electrophoresis ◽  
Pcr Amplification ◽  
Pcr Primers ◽  
Mono Lake ◽  
Ammonia Oxidizing Bacteria ◽  
Physiological Diversity ◽  
Gradient Gel Electrophoresis ◽  
Ammonia Oxidizing Bacterium ◽  
Direct Amplification ◽  
Rrna Sequences

ABSTRACT Ammonia-oxidizing bacteria were detected by PCR amplification of DNA extracted from filtered water samples throughout the water column of Mono Lake, California. Ammonia-oxidizing members of the β subdivision of the division Proteobacteria (β-subdivisionProteobacteria) were detected using previously characterized PCR primers; target sequences were detected by direct amplification in both surface water and below the chemocline. Denaturing gradient gel electrophoresis analysis indicated the presence of at least four different β-subdivision ammonia oxidizers in some samples. Subsequent sequencing of amplified 16S rDNA fragments verified the presence of sequences very similar to those of culturedNitrosomonas strains. Two separate analyses, carried out under different conditions (different reagents, locations, PCR machines, sequencers, etc.), 2 years apart, detected similar ranges of sequence diversity in these samples. It seems likely that the physiological diversity of nitrifiers exceeds the diversity of their ribosomal sequences and that these sequences represent members of theNitrosomonas europaea group that are acclimated to alkaline, high-salinity environments. Primers specific forNitrosococcus oceanus, a marine ammonia-oxidizing bacterium in the γ subdivision of the Proteobacteria, did not amplify target from any samples.

scholarly journals Direct PCR amplification from saliva sample using non-direct multiplex STR kits for forensic DNA typing

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Pankaj Shrivastava ◽  
Toshi Jain ◽  
R. K. Kumawat
Keyword(s):  
Dna Typing ◽  
Pcr Amplification ◽  
Turnaround Time ◽  
Forensic Dna ◽  
Direct Pcr ◽  
Forensic Dna Typing ◽  
Dna Profile ◽  
Pre Treatment ◽  
Dna Profiles ◽  
Direct Amplification

AbstractDue to its proficiency to provide the most discriminating results for forensic applications, medical research and anthropological studies, multiplex PCR based STR analysis has been established as the most efficient technique in the forensic DNA analysis. Several multiplex amplification kits based on 4, 5 and 6 dyes chemistry are commercially available and used in forensic DNA typing across the globe. These multiplex PCR systems are routinely used for amplification of multiple STR loci (Autosomal, Y and/or X STR’s) in the DNA extracted from various biological samples. In the routine forensic DNA testing, DNA profile obtained is compared with the DNA profile of the reference sample, which takes a certain turnaround time and employs costly lab resources. Successive development in forensic DNA typing have resulted in advent of improved multiplex kits which have reduced the effective analysis time, cost and minimized the number of steps required in comparison to conventional forensic DNA typing. Specialized direct amplification compatible multiplex kits are also available nowadays. These kits are relatively costlier but still require few pre-processing steps, which does not make them worth the hefty cost. Herein, this study, we have used non-direct multiplex STR kits to assess their efficacy for direct amplification. In the present study, 103 saliva samples were directly amplified without any pre-treatment of the samples using thirteen non-direct multiplex kits (4 dyes, 5 dyes and 6 dyes chemistry based) for forensic DNA typing. Here, we report a validated direct PCR amplification protocol from the reference saliva samples by omitting DNA extraction and quantification steps, which resulted in 80% reduction of the turnaround time. The developed protocol is cost effective, time efficient and it does not compromise with the quality of DNA profiles. To the best of our knowledge, this is the first report for direct amplification of DNA with the most commonly used non-direct multiplex STR kits without any pre-treatment of the sample. Complete DNA profiles matching all the essential quality parameters were obtained successfully from all the tested samples.

Direct amplification of reference samples with Globalfiler ® PCR Amplification Kit

2015 ◽  
Vol 5 ◽  
pp. e135-e137 ◽  
Author(s):  
N. Gouveia ◽  
P. Brito ◽  
A. Serra ◽  
F. Balsa ◽  
L. Andrade ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Direct Amplification ◽  
Reference Samples

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

1995 ◽  
Vol 74 (04) ◽  
pp. 1079-1087 ◽  
Author(s):  
Klaus-P Radtke ◽  
José A Fernández ◽  
Bruno O Villoutreix ◽  
Judith S Greengard ◽  
John H Griffin
Keyword(s):  
Rhesus Monkey ◽  
Protein C ◽  
Activated Protein C ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Heparin Binding ◽  
Reactive Center ◽  
Protein C Inhibitor ◽  
Polymerase Chain

SummarycDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey 1 liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

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