End Repair of cDNA Library
Protocol
- Mix the following components in a sterile 1.5 ml microcentrifuge tube:
Purified double-stranded cDNA: 50 μl
Nuclease-free Water: 25 μl
10X Phosphorylation Reaction Buffer: 10 μl
Deoxynucleotide Solution Mix: 4 μl
T4 DNA Polymerase: 5 μl
E. coli DNA Polymerase I, Large (Klenow) Fragment: 1 μl
T4 Polynucleotide Kinase: 5 μl
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Total volume: 100 μl - Incubate in a heat block for 30 minutes at 20°C.
- Purify end repaired DNA using a PCR column purification kit, purifying the sample on one column and eluting in 32 μl sterile water or elution buffer.
