User's Manual

16
Chapter 3: Droplet PCR (dPCR)
Preparations for dPCR
Use the Samplix Primer test PCR kit (Cat. No. RE10200) to determine the optimal primer concentration and
annealing temperature for the dPCR reaction as described in the previous chapter (Chapter 2: Droplet PCR
Reaction Design for Target Selection).
NOTE: Do not use any other reagents instead of Samplix dPCR kit (Cat. No. RE10100) for Xdrop™ dPCR droplet
production as this may compromise droplet production, droplet stability, and downstream enrichment.
Samplix provides a Positive control kit (Cat. No. CO10200) that contains template DNA and two primer pairs;
one for dPCR and one for validation qPCR after dMDA. See instructions on the next page.
Setup of dPCR reaction
1. Prepare the dPCR reaction mix following the table below.
2. Prepare mix for all reactions (40 μl reaction total per sample). Keep the reagents and the dPCR reaction
mix on ice.
3. Prepare dilutions of the correct amount of template DNA. Calculate the optimal amount of input template
DNA using the online sample calculation tool samplix.com
4. Add 2 μl of the correct amount of template DNA to 38 μl dPCR reaction mix to get 40 μl/reaction in total.
5. Allow the reaction mix to reach room temperature before loading on the dPCR cartridge followed by
immediate insertion of the dPCR cartridge on the Xdrop™ instrument.
dPCR reaction mix
1 reaction
H
2
O (molecular grade)
16,4 μl
dPCR mix (2x)
●
20 μl
dPCR Primer Forward 10 µM
0,8 μl
dPCR Primer Reverse 10 µM
0,8 μl
Total mix
38 μl
Template DNA
2 μl
Total volume
40 μl