1. General guidelines for primer design 18-30 nucleotides G/C content: 40-60% Avoid complementary sequences of primers (especially at the 3’ end) Avoid mismatches at the 3’ end Avoid 3 or more G or C at the 3’ end Avoid a 3’ end T

2. Primer design to eliminate amplification from contaminating genomic DNA

3. Primer design to detect amplification from contaminating genomic DNA

4. Primer design and analysis tools Vector NTI Primer 3 IDT SciTools PrimerQuest Oligo Analyzer

5. Real-time PCR QPCR Real-Time Vs Traditional PCR Post PCR processing Poor agrose gel resolution (semi-quantitative) Detect end point of the reaction

7. http://www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdf

12. ΔCt, ΔΔCt method to detect fold change