User manual

Droplet Digital

™

PCR Applications Guide | 73

Additional Applications

Fig. 8.1. CNV of different human samples using restriction digestion vs. undigested DNA for the same

sample for the MRGPRX1 gene.

Milepost Assay

Linkage studies can also provide information on the quality of the template, and allow you to

determine how degraded or fragmented the template might be by examining an assay that

serves as a positional anchor and a second assay located at a determined distance on the

same chromosome. This is known as a milepost assay.

Figure 8.2 represents the assay design scheme for performing a milepost assay.

Different DNA targets are designed at different lengths from an anchor assay, typically 1 kb,

10 kb, and 100 kb. Each milepost marker is run in multiplex with the anchor assay.

Copy number

Sample

Sample 1 Sample 2 Sample 3 Sample 4 Sample 5

Cut Uncut Cut Uncut Cut Uncut Cut Uncut Cut Uncut

BC_090-Rsal

BC_090

BC_106-Rsal

BC_106

BC_152-Rsal

BC_152

BC_18853-Rsal

BC_18853

BC_19108-Rsal

BC_19108

Fig. 8.2. Assay design for the milepost assay.

RPP30 anchor (chromosome 10)

(VIC)

1 kb

10 kb

100 kb

1 kb milepost marker

(FAM)

10 kb milepost marker

(FAM)

100 kb milepost marker

(FAM)