Mapping 250K/500K SNP assay

Stage 4: PCR
ABOUT THIS STAGE
chapter 4 | 96-Well Plate Protocol 61
During this stage, you will transfer equal amounts of each ligated
sample into three new 96-well plates (Figure 4.2 on page 67). You will
then prepare the PCR Master Mix, and add it to each sample.
Each plate is placed onto a thermal cycler and the 500K PCR program
is run. When the program is finished, you will check the results of this
stage by running 3 µL of each PCR product on a 2% TBE gel.
LOCATION AND DURATION
• Pre-PCR Clean Area: PCR Master Mix preparation
• PCR Staging Area: PCR set up
• Main Lab: PCR Plates placed on thermal cyclers
• Hands-on time: 1 hour
• 500K PCR thermal cycler program time: 1.5 hours; samples can be
held overnight
INPUT REQUIRED FROM PREVIOUS STAGE
The input required from Stage 3: Ligation is:
Quantity Item
1 Plate of diluted ligated samples
Keep in a cooling chamber on ice.