Protocols and Applications Guide (US Letter Size) - Promega
Protocols and Applications Guide (US Letter Size) - Promega
Protocols and Applications Guide (US Letter Size) - Promega
Create successful ePaper yourself
Turn your PDF publications into a flip-book with our unique Google optimized e-Paper software.
||||||||| 9DNA Purification<br />
Additional Resources for Fixed-Tissue Genomic DNA<br />
Isolation<br />
Technical Bulletins <strong>and</strong> Manuals<br />
TB319 MagneSil® Genomic, Fixed Tissue System<br />
Technical Bulletin<br />
(www.promega.com/tbs/tb319/tb319.html)<br />
TB382 Maxwell® 16 FFPE Tissue LEV DNA<br />
Purification Kit Technical Bulletin<br />
(www.promega.com/tbs/tb382/tb382.html)<br />
Citations<br />
Oliveira, L.R. et al. (2008) HPV infection in Brazilian oral<br />
squamous cell carcinoma patients <strong>and</strong> its correlation with<br />
clinicopathological outcomes. Mol. Med. Reports Atenas 1,<br />
123–9.<br />
To study the frequency of human papilloma virus (HPV)<br />
in Brazilian patients with oral squamous cell carcinoma<br />
(OSCC), DNA was isolated from formalin-fixed<br />
paraffin-embedded tissue samples of primary tumors <strong>and</strong><br />
their matched samples using the MagneSil® Genomic, Fixed<br />
Tissue System. Five microliters of genomic DNA was<br />
amplified using PCR Master Mix <strong>and</strong> primers for both a<br />
110bp fragment of human ß-globin gene <strong>and</strong> HPV<br />
A.<br />
B.<br />
C.<br />
<strong>Size</strong> Markers<br />
100 200 300 400 500600<br />
926-base<br />
<strong>Size</strong><br />
Marker<br />
genotype. After PCR, the product was analyzed on an 8%<br />
nondenaturing polyacrylamide gel <strong>and</strong> stained with<br />
AgNO3.<br />
972-base<br />
Amelogenin<br />
Fragment<br />
APC<br />
1.8kb<br />
Fragment<br />
Figure 9.17. Analysis of DNA purified from paraffin-embedded, formalin-fixed 10µm thin sections using the MagneSil® Genomic,<br />
Fixed Tissue System. Purified DNA was amplified, <strong>and</strong> the amplification products were analyzed on an ABI PRISM® 310 or 3100 genetic<br />
analyzer. Panel A. Amplification with a set of 16 fluorescently labeled primers. Amplification products range in size from 104 to 420 bases.<br />
Panel B. A 972-base fragment amplified using an amelogenin primer set. Panel C. A 1.8kb fragment amplified from the Adenomatosis polyposis<br />
coli (APC) gene. Increasing the extension time during amplification may help to balance yields between small <strong>and</strong> large amplification<br />
products <strong>and</strong> increase yields for large amplification products. Results will vary depending on the degree of cross-linking due to formalin<br />
fixation.<br />
<strong>Protocols</strong> & <strong>Applications</strong> <strong>Guide</strong><br />
www.promega.com<br />
rev. 3/09<br />
3931TA12_2A<br />
PROTOCOLS & APPLICATIONS GUIDE 9-26
Change language