PCR Enrich Adaptor Ligated cDNA Library (E6100)

Protocol

1. Mix the following components in a sterile PCR tube:
   Size Selected cDNA: 29 μl
   5X Phusion HF Buffer: 10 μl
   Primer 1 (25 μM): 1 μl
   Primer 2 (25 μM): 1 μl
   Deoxynucleotide Solution Mix: 1.5 μl
   Nuclease-free Water: 7 μl
   Phusion DNA Polymerase: 0.5 μl 
   -----------------------------------------------------------------
   Total volume: 50 μl
2. PCR cycling conditions:

   Cycle Step	Temperature	Time	Cycles
   Initial Denaturation	98°C	10 seconds	1
   Denaturation	98°C	10 seconds	15
   Annealing	65°C	30 seconds
   Extension	72°C	30 seconds
   Final Extension	72°C	5 minutes	1
   4°C	hold

3. Purify PCR Enriched cDNA using a PCR column purification kit, purifying the sample on one column and eluting in 30 μl sterile water or elution buffer.