Protocols and Applications Guide (US Letter Size) - Promega

| 1Nucleic Acid Amplification
Unamplified DNA
Denature and
anneal primers
Extend primers
Denature and
anneal primers
Extend primers
Denature and
anneal primers
Extend primers
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Figure 1.1. Schematic diagram of the PCR process.
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Target Region
Cycle 1
Cycle 2
Cycle 3
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Amplification of short "target" product
More (www.promega.com/pnotes/apps/pcr/)
publications
Online Tools
Amplification Product Selector (www.promega.com
/selectors/amplification/)
Citations
Bermejo-Alvarez, P. et al. (2008) Can bovine in
vitro-matured oocytes selectively process X- or Y-sorted
sperm differentially? Biol. Reprod. 79, 594–7.
To determine whether oocytes are able to select X-bearing
or Y-bearing spermatozoa, the authors performed in vitro
fertilization of bovine oocytes with X-sorted semen,
Y-sorted semen, a mixture of X- and Y-sorted semen, and
unsorted semen. The gender of the resulting embryos was
determined by amplifying two DNA targets: a Y
chromosome-specific target for gender assignment and a
bovine-specific satellite sequence as a control. PCRs were
Protocols & Applications Guide
www.promega.com
rev. 12/09
Cycles 4–30
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=Short "target" product
=Long product
performed using GoTaq® Flexi DNA Polymerase (1 unit
per 25μl reaction), and amplified products were analyzed
by agarose gel electrophoresis followed by ethidium
bromide staining.
PubMed Number: 18579751
Staniszewska, I. et al. (2008) Integrin alpha9 beta1 is a
receptor for nerve growth factor and other neurotrophins.
J. Cell Sci 121, 504–13.
The authors investigated the ability of α9β1 integrin to act
as a neurotrophin receptor and affect cell signaling
pathways. As part of the study, RT-PCR was used to detect
the presence of other neurotrophin receptors in their model
cell line SW480. Reverse transcription was performed using
the Reverse Transcription System and 1μg of total RNA
isolated using the SV Total RNA Isolation System. The
resulting cDNA (5μg) was amplified for 35 cycles (β-actin
1160MA06_5A
PROTOCOLS & APPLICATIONS GUIDE 1-2