eSeminars
&
eConference & Webinars
central topics = qPCR
& dPCR &
RNAi & Molecular-Biology
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This
streaming portal is dedicated to
scientists from the community of
qPCR, digital PCR, Next Generation
Sequencing (NGS), MicroGenomics (MG)
and Molecular Diagnostics (MDx).
You’ll find here all the records
from around 500
presentations held at qPCR
& NGS and MG Events in the past
years – qPCR 2010 in Vienna
to
GQ2023
in Freising-Weihenstephan, and
MicroGenomics 2014 in Paris.
We provide the presentations via
movie streaming technology in high
quality – high resolution and
perfect sound quality in high
speed – on any internet
browser or mobile device,
including Android or iOS.
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iBiology
iBiology’s mission is to
convey, in the form of open-access
free videos, the excitement of
modern biology and the process by
which scientific discoveries are
made. Our aim is to let you meet the
leading scientists in biology, so
that you can find out how they think
about scientific questions and
conduct their research, and can get
a sense of their personalities,
opinions, and perspectives. We also
seek to support educators who want
to incorporate materials that
illustrate the process and practice
of science into their curriculum.
This project is made possible by the
good will of many biologists who are
committed to making their work
broadly accessible and to conveying
the excitement of biology to a
worldwide audience.
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www.iBiology.org
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Identification
of
MRSA using 3-colour Crystal digital
PCR
Tuesday, October 24, 2017
8 AM PDT | 11
AM EDT | 4 PM
BST | 5 PM CEST |
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Methicillin
resistant
Staphylococcus aureus (MRSA) is a
major pathogen associated with
complicating hospital and community
medicine. Drug resistance increases
the risk of complications and can
challenge routine procedures such as
surgery. As different species, such as
Staphylococcus epidermidis, can also
carry resistance, this can further
increase the challenge of diagnosis
using culture and/or molecular
methods.
This webcast describes a digital PCR
approach that uses a three colour
detection platform (Naica Crystal
Digital PCR System, Stilla
Technologies) to count the number of
genes that confer resistance and
compare this with the number of genes
from S. aureus and S. epidermidis
within a sample. This method may be
useful to determine which organism is
carrying the resistance gene.
During this webcast you will:
- Understand 3-colour
multiplexing digital PCR
- Understand The Naica System
Workflow
- Learn a new method to
determine resistance gene and drug
resistance
- Given a practical study :
identification of MRSA with the
Naica System
- Learn about digital PCR for
infectious diseases applications
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Nature
-
Podcast Podcast
-
index page
Nature
- Video Streaming
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Science
- Podcast
Science
- Multimedia
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Animations
of Inhibitory RNA Action:
- Nature Reviews
Genetics - Nature Reviews
Genetics (2001)
"Post-Transcriptional Gene
Silencing by Double-Stranded RNA"
- Nature Reviews
- A high quality movie describing
inhibitory RNA events and
mechanisms
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qPCR
talks around the world:
Reliable
Quantification
by qPCR
a talk by Steve Bustin
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RNA
Quantification in Research and
Diagnostics
a talk by Mikael Kubista
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The Importance of Controls and Novel
Solutions for Successful Real-time
qPCR
a talk by Qiagen
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Learn Why You Might Be One of the 90%
Using Unsuitable qPCR Reference Genes
a talk by Bio-Rad
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Six Tips for Increasing the
Reproducibility of qPCR Experiments
a talk by Bio-Rad
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How to Minimize Contamination in qPCR
Experiments
a talk by Bio-Rad
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Quantitative PCR
by Matthias Dobbelstein
Molecular Oncology, GZMB,
Universitätsmedizin Göttingen, Germany
9
videos on YouTube
Quantitative PCR. This series of
short chalk-talk videos will cover the
principles and calculations associated
with quantitative PCR. The seminar is
also part of a methods course provided
to PhD students of the Göttingen
Graduate School of Neurosciences and
Molecular Biosciences, GGNB.
- Quantitative PCR -
Introduction A 2:35
- Quantitative PCR -
Introduction B 14:37
- Quantitative PCR -
real-time PCR and SybrGreen
fluorescence 14:05
- Quantitative PCR - the
deltaCt method 9:27
- Quantitative PCR - deltaCt
in the real world 10:21
- Quantitative PCR -
RT-PCR 7:36
- Quantitative PCR - internal
controls 8:54
- Quantitative PCR - the
deltadeltaCt method 6:22
- Quantitative PCR - the
melting curve 11:25
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Molecular
Biology On-Demand Webinars:
- Direct PCR: Amplify Without
Purification
- PCR Reaction Setup and
Cycling
- Optimize Your PCR
- The Essentials of Nucleic
Acid Sample Preparation
- How to Avoid Common
Mistakes in the qPCR Workflow
- Your Key to Success in NGS:
The Library Preparation Workflow
- DNA Polymerases at Work:
Finding Your PCR Champion
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Molecular
Biology On-Demand Webinars:
- Restriction Enzyme
Digestion: Cutting DNA the Right
Way
- Optimize Your Cloning: No
More Trouble With Ligation
- The Essentials of Real-Time
PCR
- Cloning Workflow
Considerations
- Direct PCR
- Thermo Scientific Direct
PCR Kits and Applications
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Nanostring Webinars
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WEBINAR
-- Characterizing the Performance of
qPCR Instruments – Approaches for
Assessment and Comparison
The
breadth of instruments available for
quantitative PCR (qPCR) has
continued to grow in the past 5-10
years. With older platforms now
being retired and an abundance of
new technologies available to
replace them, lab managers,
technicians, and researchers will
need to effectively compare and
evaluate the performance of these
platforms. While new features such
as multiplexing, microfluidics, and
integration with liquid handling
automation have enabled higher
throughput and lower operating
costs, it has made it increasing
complex to readily compare different
types of instruments and their
respective performance
characteristics.
As
the list of features and
specifications grows, understanding
some of the key metrics of
instrument performance will become
critical for evaluating platforms
that will best meet the needs of a
laboratory’s application focus and
assay requirements. Unfortunately,
instrument vendors have not
consistently conformed to any
particular standards for defining
and assessing performance
characteristics of qPCR instruments
and rarely have the methods been
adequately documented in the product
literature.
In
this webinar, we will define several
key performance metrics of qPCR
instruments such as dynamic range,
Cq uniformity, sensitivity, and
resolution, and further discuss
their importance in practical terms.
Using data from characterization and
verification studies performed on
the IntelliQube instrument from
Douglas Scientific, we will also
review approaches to evaluating
these metrics, including assays and
software tools that streamline the
analysis and interpretation of
performance testing results.
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Real-time
PCR
tutorial | qPCR that Works
Scientists worldwide trust real-time
PCR reagents from Takara Bio–even
for the most demanding qPCR
experiments.
When
you
use real-time PCR reagents that are
sensitive and specific, you can spend
less time on PCR troubleshooting and
more time on moving your research
forward. Takara Bio offers qPCR kits
for use with both SYBR Green detection
and TaqMan probes.
Part
1:
Getting You Started
- Find
the right detection method
- Study
your gene expression by RT-qPCR
- Check
the accuracy of your results
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Speaker:
Professor
Mikael Kubista
TATAA
Biocenter
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Compatible
with
all major real time PCR instrument
systems, these products allow you to
obtain accurate, consistent results
from a wide variety of sample types,
even when other reagents fail. Want to
try Takara Bio real time PCR reagents
for yourself? Samples are available
for many of our qPCR reagents.
Part 2: Interpreting Your Data
- Determine
Cq
values
- Obtain
absolute quantification of your
starting DNA
- Define
a good reference for relative
quantification
- Follow
MIQE guidelines
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Speaker:
Robert
Sjöback,
Ph.D.
TATAA
Biocenter
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Count
on Takara Bio to help you choose the
best qPCR products for plant, soil,
blood, paraffin-embedded tissues,
archival or degraded samples or when
conducting gene expression studies,
forensic research analyses, or other
specific applications. Want to learn
more about real-time PCR? Three videos
produced by TATAA Biocenter in Europe
help you understand everything from
the basics of qPCR to data analysis
techniques. Each 20- to 30-minute
video can be watched at your
convenience from the comfort of your
office, lab, or home.
Part 3: Troubleshooting Your
Results
- Identify
potential
pitfalls in qPCR data
- Detect
PCR inhibition
- Control
the quality of RNA and reverse
transcription
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Speaker:
Kristina
Lind,
Ph.D.
TATAA
Biocenter
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Online
Real-Time PCR Training
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Module
1:
Fundamentals
of real-time PCR |
Module
2:
Assay
design choices |
Module
3:
Normalization |
Module
4:
Multiplexing |
Module
5:
Reagent
choices |
Module
6:
Theory of analysis |
| Access
an overview of real-time PCR
instrument choices and learn how
real-time PCR differs from endpoint
PCR. How can you use real-time PCR
in your research? What is the
difference between TaqMan® chemistry
and SYBR® Green chemistry? |
What
should you consider when planning
your gene expression assays? What is
the impact of multitranscript assays
and assay specificity? How do you
design custom assays? |
What
is normalization and why is it
important? What are the types of
normalizers and what are their
functions? How does multigene
normalization work? |
What
is multiplexing, and when and how do
you use it? |
Discussion
topics
include setting up your real-time
PCR reaction, and preparing master
mixes and reagents for plate
additions. |
Perform
dynamic
range testing with standard curves,
quantify your data using the ΔΔCt
method, and gain a better
understanding of absolute vs.
relative quantification. |
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View
trainings
here |
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Whether you’re new to PCR or need
more in-depth learning guides our
video library will have all the
resources you need. See our
Veriti® Thermal Cycler in action,
catch up with Ph.Diddy and Ph.Diva
and go behind the scenes. New videos
will be added periodically so be
sure to bookmark this page for
future reference.
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Introduction
to
Real-Time PCR
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Introduction
to
TaqMan® Copy Number Assays
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Introduction
to
TaqMan® Gene Expression Assays
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Introduction
to
TaqMan® Protein Assays
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Fundamentals of Real-Time PCR |
Introduction to Digital PCR |
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To
view
these recorded webinars,
please ensure that your pop-up
blocker is disabled !
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qPCR and MIQE
Seminar Series
"MIQE Oldies but Goodles" from the Sigma
Aldrich Learning Center
As part of our customer education
program, we have provided two recorded
seminar series covering the topics of qPCR
and MIQE. The recorded sessions are intended
to provide a high level overview of these
subject matters. We have kept the lessons
concise so that you can enjoy a self-paced
learning program.
| Seminar
Title |
Presenter |
Recording
Length
(hours : minutes : seconds) |
| Primer and Probe Design |
Ashley Heath, PhD |
0:06:32 |
| An Introduction to qPCR
Concepts |
Mudassir Mohammed, PhD |
0:09:37 |
| Selecting a qPCR Basic
Detection Chemistry |
Mudassir Mohammed, PhD |
0:12:35 |
| Choosing a Fluorophore /
Quencher Combination |
Anders Bergkvist, PhD |
0:11:30 |
| Chemistries for More
Challenging qPCR Assays |
Mudassir Mohammed, PhD |
0:15:18 |
| MIQE Concepts |
Marina Wiklander, PhD |
0:03:19 |
| Reference Gene Validation |
Anders Bergkvist, PhD |
0:12:47 |
| Data Analysis Guidelines |
Anders Bergkvist, PhD |
0:10:42 |
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| Seminar
Title |
Presenter |
Recording
Length
(hours : minutes : seconds) |
| MIQE: Assay Design
Considerations |
Tania Nolan, PhD |
0:17:37 |
| MIQE: Sample Derived
Inhibitors |
Tania Nolan, PhD |
0:13:04 |
| MIQE: RNA Quality
Considerations |
Tania Nolan, PhD |
0:15:31 |
| MIQE: RNA Quantity and RT
Considerations |
Tania Nolan, PhD |
0:16:38 |
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RNA Integrity
Database (RINdb)- Bioanalyzer RNA Profiles
Marc Valer, Microfluidics Program
Manager, Genomics, Agilent Technologies,
Santa Clara.
Synopsis:
Thousands
of users today trust the bioanalyzer 2100
for qualifying total RNA samples, looking
for integrity, purity, recovery, and
consistency information for sample
preparation methods, assisting them to
determine the optimal conditions for their
experimental design. Marc Valer describes
the use of Agilent's new RNA Integrity
Database (RINdb) to assist in the
development of experimental protocols,
particularly sample preparations, by
providing a means for researchers to
contextualize RNA profiles.
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Qiagen Webinars
Register for a live Webinar and hear a
live talk about the cutting-edge
technologies of your choice followed
by a Q&A session. The speaker will
answer as many of your questions as
possible during the session. Any
remaining questions will be answered
by personal e-mail after the Webinar.
Alternatively, you can listen to one
of our previously recorded Webinars.
http://www1.qiagen.com/events/webseminars/default.aspx
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- miRNA purification and
detection — new tools in
expression profiling and biomarker
development
- Recent progress in RNAi
screening
- A successful and affordable
RNAi solution for every lab
- Novartis scientists share
their experiences in multiplex,
real-time PCR
- Transcriptome-wide miRNA
quantification by RT-PCR
- Accelerate your PCR and
qPCR
- and much
more..........................
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PODCAST: The
Introduction to Molecular
and Cellular Biology lectures
include text, images, and audio. Each
lecture webpage is synchronized to the audio
component. In addition, the lectures are
available as a podcast subscription.
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Fast
PCR and real-time PCR tutorials
(by Bio-Rad)
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The
presentations listed below provide
background and instruction about specific
concepts in PCR.
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Free of
charge web seminars
Sigma-Aldrich is pleased to be
able to bring you a series of webex seminars dedicated to
qPCR. They will include technical
presentations from international speakers
discussing the latest techniques and aspects
of qPCR. To register, go
to
QPCR Seminars
Sigma-Aldrich has the pleasure of
inviting you to a seminar series devoted to
the technical aspects of qPCR and RT-qPCR.
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Available
Seminars: |
| Author |
Seminar
Title |
| Mudassir
Mohammed, Ph.D. |
An
Introduction to qPCR |
| Valerie
Hawkes, Ph.D. |
Assay Design
and Optimisation |
| Neven Zoric,
M.Sc. |
Approaches to
Normal qPCR — Facing the Challenge
of Normalization |
| Tania Nolan,
Ph.D. |
Deriving a
Troubleshooting Protocol |
| Mikael
Kubista, Prof. |
A Statistical
Approach to qPCR Gene Expression
Data Analysis |
| Marina
Tshuikina, Ph.D. |
An
Introduction to Epigenetics Analysis
by qPCR |
| Valerie
Hawkes, Ph.D. |
Improving the
Discrimination and Sensitivity of
qPCR Assays using LNA |
| Chris Bass,
Ph.D. |
Development of
DNA-based Diagnostic Assays for
Sensitive Screening of Mosquito
Disease Vector Populations |
| Michael
Pfaffl, Ph.D. |
Influence of
RNA Integrity on Real-Time RT-PCR
Quantification Data |
| Vladimir Benes |
microRNA
Profiling |
| Rebecca Hands |
Technical Tips
on LCM Extraction for mRNA Profiling |
| Jens Stolte |
cDNA
Amplification with GenomePlex®
Complete Whole Genome Amplification
Kit |
| Tanya Novak,
M.Sc. |
The SPUD Assay
Has an Important Role in the
Interpretation of Detecting
Pneumocystis DNA in Clinical
Specimens |
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Transcriptional
Regulation of Eukaryotic Genomes
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Fundamentals
of
Real-Time PCR
(by
Applied
Biosystems,
47 minutes)
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The Eppendorf real-plex system
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Audio Slide Show: SmartCycler®
System for qPCR
(from
Cepheid)
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RNA
integrity Audio slide shows
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The OpenArray™ Nanotiter Plate Technology
and Applications
Understanding biological
complexity arising from patterns of gene
expression requires accurate and precise
measurement of RNA levels across large
numbers of genes simultaneously. The
preferred method for quantitative
transcriptional analysis is real time PCR
(rt-PCR) in a 96- or 384-well microtiter
plate. Scaling rt-PCR to higher throughputs
is intrinsically limited by cost and
logistic considerations. Alternatively
hybridization
microarrays are capable of measuring
transcription of many thousands of genes
simultaneously yet are limited by low
sensitivity, accuracy and sample throughput.
We demonstrate a hybrid
approach by combining the superior accuracy,
precision and dynamic range of rt-PCR with
the high density parallelism of a microarray
in an array of 3,072 real-time, 33 nL
polymerase chain reactions the size of a
microscope slide. Real-time
PCR in this nanotiter plate format results
in substantial savings in reagents,
measurement time and productivity. We
demonstrate system performance by measuring
tissue-specific expression of kinase genes
in human heart and liver samples and
transcriptional modulation by small-molecule
perturbation of the TNF-alpha signaling
pathway in HUVEC cells. Compared
with the same assay in a 384-well
microplate, we measured equivalent rt-PCR
performance with a 64-fold reduction in
assay volume, a 24-fold increase in
analytical throughput and a workflow
compatible with standard microplate
protocols in an easy-to-use fomat.
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Dynamic Arrays to Measure Expression of
Nucleic Acids and Proteins
Michael
Lucero of Fluidigm speaking at AMT 2006
Click
here
to launch presentation
A
dynamic array is a biochip that employs
integrated channels and valves in a matrix
architecture. This matrix architecture is
a breakthrough in efficiency because
performing the same set of reactions by
hand or with a robot would require orders
of magnitude more sample, reagents, and
pipetting steps.Dynamic
arrays have been introduced that handle
homogeneous or heterogeneous assays, such
as real-time qPCR and ELISAs,
respectively. BioMark™ dynamic arrays for
qPCR accept 48 samples and 48 primer/probe
sets. The components are combined into
2,304 assays (48 x 48).The
chip is ideal to validate expression for
48 genes on samples from many individuals.
Projected throughput of future chips is
~10,000 reactions. BioMark™ dynamic arrays
for immunoassays accept 48 samples and 18
antibody pairs and generate 1,728 assays.
Integrated valves prevent mixing between
antibody pairs.Thus,
dynamic arrays prevent signal crosstalk
and eliminate the need to optimize
antibody pairs for multiplexing in one
vessel, a requirement with other formats.
Instrumentation automates the loading of
chips and analysis of results. Data
produced on BioMark™ dynamic arrays for
both applications will be presented,
demonstrating a sensitivity of detection
equivalent to conventional microwell
plates while generating orders of
magnitude higher throughput.
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A Sequence Oriented Comparison of Gene
Expression Measurements Across Different
Hybridization-Based Technologies
Winston
Kuo of Harvard School of Dental Medicine
speaking at AMT 2006
Click
here
to launch presentation
Gene
expression microarrays have made a
profound impact in biomedical research.
The diversity of platforms and analytical
methods has made comparison of data from
multiple platforms very challenging. The
presentation will describe a framework for
comparisons across platforms and
laboratories, where probe sequences were
utilized from each vendor to map of genes
across platforms.
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High-throughput Measurement of Expression
Signatures Using Dynamic Arrays for qPCR and
Immunoassays
Michael
Lucero of Fluidigm speaking at the European
Biomarkers Summit 2006
To purchase a DVD of all the
presentations featured at the European
Biomarkers Summit, please go to the Select
Biosciences website.
Click here to
launch presentation
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RNAi screening – advanced tools to
accelerate translational research and drug
discovery
- In
recent
years, RNAi screening using
synthetic siRNA libraries has
become a popular tool for
elucidating gene functional
pathways and for target
identification. Find out about
the essentials of RNAi screening
and the latest tools developed
by QIAGEN to enable its broad
application from Dr. Eric Lader,
QIAGEN’s Associate Director of
RNAi research.
- RNAi screening - advanced tools
to accelerate translational research and
drug discovery.
- Listen to this exciting Webinar
now.
- RNAi technology and genome-wide
expression profiling - assessment of
specificity and pathway analysis.
- Listen to this exciting Webinar
now.
- Qiagen
Webinar
web page
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Identification of New miRNA Markers for
Breast Cancer with LNA Microarrays
Thomas
Litman, Exiqon, speaking at RNAi Europe 2006
Click here to
launch presentation
Abnormal
expression
of
microRNAs (miRNAs) in cancer implies that
these small ~22-nucleotide molecules play
a role in oncogenesis, and therefore may
comprise a novel class of diagnostic and
prognostic signatures. Here, we are
studying the global expression profiles of
miRNAs in breast cancer and adjacent
non-malignant breast tissue in order to
identify possible new biomarkers for
breast cancer
Biopsies
from primary tumors and from the proximal
tissue (1 cm and 5 cm from the border zone
of tumor) were collected from female
patients (age 55-69) undergoing surgery
for invasive ductal carcinoma. Total RNA
was extracted and analyzed for global
miRNA expression on a miRCURYTM LNA-based
microarray platform containing capture
probes for over 400 miRNAs.
Our
analysis of miRNA expression patterns from
tumor and proximity tissue revealed
numerous differentially expressed miRNAs,
including those reported to be associated
with breast cancer, such as let-7a/d/f,
miR-125a/b, miR-21, miR-32, and miR-136.
In
addition, we have identified several
miRNAs that have not previously been
connected with breast cancer. Some of
these novel miRNA signatures could have
diagnostic and prognostic potential for
breast cancer patients.
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RNAi Based Toold in Apptosis Research
Yu Shen,
Abbott Laboratories, speaking at RNAi Europe
2006
Click here to
launch presentation
Several
cases of off-target gene silencing were
identified in our siRNA library screens
(Lin X et.al. Nucleic Acids Research 33:
4527-35, 2005). However, despite the
complications of off-target gene
silencing, we successfully identified
three cancer targets by screening an siRNA
library against the “druggable genome”
using a cell-death assay.
In
addition, in an siRNA-based synthetic
lethal screen, we found that knockdown of
survivin led to the selective killing of
K-Ras mutant cells. We also explored
RNAi-based methodology for target
validation in animal models.
We
developed a tightly regulated shRNA
expression system (Lin X. et.al. FEBS
letter, 577: 376-80, 2004) and used this
system to prepare stable tumor cell lines
that conditionally expressed an shRNA for
HIF-1a. These tumor cells were implanted
in mice to form tumors that served as a
versatile tool for studying the effects of
inhibiting HIF-1 in vivo (Li L et.al.
Cancer Research, 65: 7249-58, 2005).
Finally,
we investigated several methods for the
creation of germline knockdown animals. By
modifying the standard methods, we
successfully increased the transgenic
frequency and F1 transmission rate and
created tyrosinase knockdown mice with a
lighter-coat-color phenotype that can be
stably transmitted to the next generation.
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OpenGenomics Peer Science
http://www.opengenomics.com/
Welcome
to the OpenGenomics Peer Science series,
where your colleagues discuss their latest
findings in Genomics research. With
technical webcasts from your peers,
podcasts providing distilled takes on
research breakthroughs, and the latest in
published papers, Open Genomics is
dedicated to providing fresh perspectives
on pressing research questions. Check here
regularly for the latest developments in
Genomics.
Published
Papers
A
searchable database of the latest
publications using Agilent's DNA
microarray platform. Search by
application, date, or keyword.
Webcasts
Each
webcast contains a short scientific talk
on a specific area of research, presented
by the researcher. These webcasts are
15-20 minutes long and are available on
demand.
Podcasts
Each
Podcast is developed as an informational
brief highlighting innovative approaches
to fundamental research questions,
available as an RSS feed to your IPod®, or
available for listening on your computer.
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Weekly Podcasts from the GENcast
Network
http://www.genengnews.com/gencasts.aspx
These
weekly podcasts feature interviews with
leading biotech researchers, newsmakers,
and thought leaders. Topics revolve around
the critical scientific and business
issues that impact the global
biotechnology industry, beginning with new
discoveries in the lab and then moving
onto R & D, biomanufacturing, and
final product commercialization. Trends,
novel technologies, opinions, recent
developments, how-to advice, and much more
are discussed in our podcasts in a lively,
to-the-point, and informative style. New
every Thursday, you can listen right from
the GEN website or you can subscribe using
the button below to have it download each
week automatically to your iPod or mp3
player!
- THE
INVENTOR OF PCR - GEN's
Editor-in-Chief John Sterling interviews
the Nobel Prize winning inventor of PCR,
Dr. Kary Mullis. (2/15/2007) sponsored
by: Eppendorf
- RNAi TECHNOLOGY -
Richard Jorgensen, Ph.D., from the
University of Arizona (3/22/2007)
sponsored by: Sigma-Aldrich
- qPCR
AND LATE-PCR, AN ADVANCED TECHNIQUE
FOR DNA AMPLIFICATION -
Lawrence Wangh, Ph.D., from Brandeis
University (2/22/2007) sponsored
by: Eppendorf
- PROBE-BASED,
REAL-TIME
QUANTITATIVE
PCR ASSAY DESIGN AND APPLICATIONS
- Gregory L. Shipley, Ph.D.,
Director, Quantitative Genomics Core
Laboratory, The University of Texas
Health Science Center-Houston.
(2/1/2007) sponsored by: Eppendorf
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for Windows (.wmv)
for Macintosh (.mov)
for
Printing (.pdf)