Chapter 08

Author: Kelly Elkins

© 2013 Elsevier, Inc. All rights reserved.

© 2013 Elsevier, Inc. All rights reserved.

Figure 8.1 PCR primer locations (bold) to be used to amplify Homo sapiens thyroid peroxidase (TPO) on chromosome 2, NCBI Nucleotide Sequence (Accession: NG_011581) (64 bp). The sequence is shown from 5’ to 3’.

© 2013 Elsevier, Inc. All rights reserved.

Figure 8.2 Exponential amplification curves that show sample results of forty cycles of real time PCR with 1 ng K562 DNA template. Curves of K562 DNA as detected by SYBR green I.

© 2013 Elsevier, Inc. All rights reserved.

Figure 8.3 K562 human DNA standard curve: CT vs. log DNA concentration.

© 2013 Elsevier, Inc. All rights reserved.

Figure 8.4. A. Sample melt curve plot from 50-95 °C showing the temperature inflection at 81 °C for the K562 DNA shown in Figure 8.2 (left). B. First derivative plot of the melting curve from 50-95 °C showing the predominant peak at 81 °C for the K562 DNA (right).

© 2013 Elsevier, Inc. All rights reserved.

Figure 8.5 Sample 1% agarose gel of PCR of K562 DNA using TPOX primers. Lanes 1 contains the DNA Logic ladder (sizes indicated), Lanes 3, 7, 8, 10, and 11 contain the 64 bp amplicon.

© 2013 Elsevier, Inc. All rights reserved.

Figure 8.6 Setup of cycling and temperature and plate protocols on BioRad iQ5.