Biomedical Engineering Reference
In-Depth Information
Fig. 1. Four different-sized DNA molecules are shown in the upper diagram. The
arrow pointing to the right represents the forward primer and the arrows pointing to
the left side represent different reverse primers. The DNA molecules can be amplified
and detected by the real-time polymerase chain reaction only if both the forward and
reverse primers can be annealed to the DNA molecules. With the increase of amplicon
size, the number of DNA molecules detected by the system is decreased.
encoded RNAs (EBER) were developed
(
11
)
. The amplicon lengths of these
PCR assays ranged from 82 to 1000 bp. These quantitative real-time assays
could only detect DNA molecules longer than their respective amplicons.
Therefore, for the same sample, the EBV DNA concentration measured by an
assay with longer amplicon would be lower than that measured by an assay
2. Materials
2.1. Sample Collection
Ethylenediamine tetraacetic acid (EDTA)-containing blood collection tubes
for plasma collection.
2.2. DNA Extraction
1.
Absolute ethanol.
2.
QIAamp Blood Midi and Mini Kits (Qiagen, Hilden, Germany).
a. Protease.
b. Lysis buffer.
