REFERENCIAS
- F. Luo, Z. Li and G. Dai et al., Simultaneous detection of different bacteria by microchip electrophoresis combined with universal primer-duplex polymerase chain reaction, Journal of Chromatography A, https://doi.org/10.1016/j.chroma.2019. 460734
- Octava edición Biología Molecular, Lina María Martínez Sánchez, Natalia Vargas Grisales, Laura Mejía Cardona, Felipe Osorio Ospina, Sergio Ramirez Pulgarín.
Hotspot mutation and fusion transcript detection from the same non-small cell...Thermo Fisher Scientific
The presence of certain chromosomal Header
rearrangements and the subsequent fusion
gene derived from translocations has been
implicated in a number of cancers. Hundreds of
translocations have been described in the
literature recently but the need to efficiently
detect and further characterize these
chromosomal translocations is growing
exponentially. The two main methods to identify
and monitor translocations, fluorescent in situ
hybridization (FISH) and comparative genomic
hybridization (CGH) are challenging, labor
intensive, the information obtained is limited,
and sensitivity is rather low. Common sample
types for these analyses are biopsies or small
tumors, which are very limited in material
making the downstream measurement of more
than one analyte rather difficult; obtaining
another biopsy, using a different section or
splitting the sample can raise issues of tumor
heterogeneity. The ability to study mutation
status as well as measuring fusion transcript
expression from the same sample is powerful
because you’re maximizing the information
obtained from a single precious sample and
eliminating any sample to sample variation.
Here we describe the efficient isolation of two
valuable analytes, RNA and DNA, from the
same starting sample without splitting, followed
by versatile and informative downstream
analysis. This methodology has been applied to
FFPE and degraded samples as well as fresh
tissues, cells and blood. DNA and RNA were
recovered from the same non-small cell lung
adenocarcinoma sample and both mutation
analysis, as well as fusion transcript detection
was performed using the Ion Torrent PGM™
platform on the same Ion 318™ chip. Using
10ng of DNA and 10ng of RNA input, we
applied the Ion AmpliSeq™ Colon and Lung
Cancer panel to analyze over 500 COSMIC
mutations in 22 genes and the Ion AmpliSeq™
RNA Lung Fusion panel to detect 40 different
fusion transcripts.
Hotspot mutation and fusion transcript detection from the same non-small cell...Thermo Fisher Scientific
The presence of certain chromosomal Header
rearrangements and the subsequent fusion
gene derived from translocations has been
implicated in a number of cancers. Hundreds of
translocations have been described in the
literature recently but the need to efficiently
detect and further characterize these
chromosomal translocations is growing
exponentially. The two main methods to identify
and monitor translocations, fluorescent in situ
hybridization (FISH) and comparative genomic
hybridization (CGH) are challenging, labor
intensive, the information obtained is limited,
and sensitivity is rather low. Common sample
types for these analyses are biopsies or small
tumors, which are very limited in material
making the downstream measurement of more
than one analyte rather difficult; obtaining
another biopsy, using a different section or
splitting the sample can raise issues of tumor
heterogeneity. The ability to study mutation
status as well as measuring fusion transcript
expression from the same sample is powerful
because you’re maximizing the information
obtained from a single precious sample and
eliminating any sample to sample variation.
Here we describe the efficient isolation of two
valuable analytes, RNA and DNA, from the
same starting sample without splitting, followed
by versatile and informative downstream
analysis. This methodology has been applied to
FFPE and degraded samples as well as fresh
tissues, cells and blood. DNA and RNA were
recovered from the same non-small cell lung
adenocarcinoma sample and both mutation
analysis, as well as fusion transcript detection
was performed using the Ion Torrent PGM™
platform on the same Ion 318™ chip. Using
10ng of DNA and 10ng of RNA input, we
applied the Ion AmpliSeq™ Colon and Lung
Cancer panel to analyze over 500 COSMIC
mutations in 22 genes and the Ion AmpliSeq™
RNA Lung Fusion panel to detect 40 different
fusion transcripts.
SLIDE CONTAIN BREIF NOTE ON PCR. IT CONTAINS 21 SLIDES INCLUDING, WHAT IS PCR? COMPONENTS, WORKING MECHANISM, APPLICATIONS, CONCLUSION, AND SOME REFRENCES, HISTORY ALSO
this ppt contain about pcr technique and its three process,primers in pcr,dna polymerase in pcr,melting temp of dna in pcr and applications of pcr technology
The study of the complete set of RNAs (transcriptome) encoded by the genome of a specific cell or organism at a specific time or under a specific set of conditions is called Transcriptomics.
Transcriptomics aims:
I. To catalogue all species of transcripts, including mRNAs, noncoding RNAs and small RNAs.
II. To determine the transcriptional structure of genes, in terms of their start sites, 5′ and 3′ ends, splicing patterns and other post-transcriptional modifications.
III. To quantify the changing expression levels of each transcript during development and under different conditions.
Francesca Gottschalk - How can education support child empowerment.pptxEduSkills OECD
Francesca Gottschalk from the OECD’s Centre for Educational Research and Innovation presents at the Ask an Expert Webinar: How can education support child empowerment?
Introduction to AI for Nonprofits with Tapp NetworkTechSoup
Dive into the world of AI! Experts Jon Hill and Tareq Monaur will guide you through AI's role in enhancing nonprofit websites and basic marketing strategies, making it easy to understand and apply.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
A Strategic Approach: GenAI in EducationPeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
1. Sofía Adarve Rengifo
III semestre
Facultad de Medicina
Universidad Pontificia Bolivariana
2020 – 1
2. INTRODUCTION
PCR (Polymerase Chain Reaction) MCE (Microchip Electrophoresis)
This method amplifies small samples of
genes, DNA or RNA segments that are
present in different mixtures.
It is use (among others) for DNA
manipulation and identification of virus and
bacterias
This technique uses an electric field in order
to separate charge molecules such as
proteins and nucleic acids.
It is widely used because of its fast analysis
speed and its minimum samples
requirements
4. OBJECTIVE
This study aimed to simultaneously
detect two kinds of bacteria by
microchip electrophoresis
combined with universal primer-
duplex polymerase chain reaction
Salmonellaenterica
serovarS.Typhimurium
Pseudomonas
aeruginosa
6. Es el tipo de MCE mas usado por su alta sensibilidad
MCE – LIF
(laser induced fluorescence)
MATERIALES Y MÉTODOS
Se usó para identificar y
diferenciar C1 y C2
Las moléculas se separan en el campo
eléctrico de acuerdo a su carga y peso
molecular
Las moléculas migran al electrodo
opuesto según su carga
Investigación
7. MATERIALES Y MÉTODOS
Primer dupplex
PCR
Amplificación
enzimática in
vitro
A partir de una pequeña muestra
de un segmento de ADN se
obtienen múltiples copias de la
secuencia.
Se necesita:
- Una sola hebra de ADN
- Un primer
- Una DNA polimerasa
Se utilizó para detectar las
concentraciones de P. aeruginosa y
S. Typhimurium por medio de la
cuantificación de C1 y C2
Investigación
Permite amplificar simultáneamente
diferentes secuencias
10. DISCUSSION
AUTHOR CONTRIBUTION YES OR NOT
Y. Zhang, et al.
“Under optimal conditions, the detection of S.
Typhimurium by MCE-LIF was achieved within
135 s with a limit of detection (S/N = 3) of
3.37 × 102 CFU mL−1 ”
K.Y. Wang, Y.L. Zeng, X.Y. Yang,
W.B. Li, X.P. Lan
“After centrifugation, the supernatant was
used as a template for the amplification of
bound aptamers by PCR with FITC- or trB-
labeled primers (20–30 cycles of 0.5 min at
94°C, 0.5 min at 52°C, 0.5 min at 72°C, followed
by 5 min at 72°C)."
Y. Zhang, et al.
“The limits of detection for the three bacteria
were ranged from 40 to 70 CFU mL−1.”
11. CONCLUSIONS
Parameters as primer concentration,
MgCl2 concentration and temperature
need to be taken into consideration in
order to develop a sensitive and specific
method of bacteria detection.
The idea of using the method exposed in
this article in the daily medical practice is
tempting, due to the fact that S.
Thymurium is associated with gut diseases
in humans and P. aeruginosa is related
with respiratory problems.
Mixing PCR method with MCE
technique have positive effects in
bacteria determination. When working
together, the LOD (lower limit of
detection) decreases notoriously, which
means that the techniques have
become more sensitive to the colonies
forming units, making the time of
detection shorter.