This page was created as an assignment for an undergraduate course at Davidson College.

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Primers used to clone the 5 isocitrate dehydrogenase genes

Ideally all primers should have the following characteristics: (Campbell, 2002)

1. A length between 18-30 nucleotides

2. A Tm between 44-46 Celsius

            Melting Temperatures calculated at this site:

http://members.aol.com/_ht_a/lucatoldo/myhomepage/JaMBW/3/1/9/index.html

3. At GC content between 40-60%

Reverse complement of gene for reverse primers was obtained at this site:

http://www.ualberta.ca/~stothard/javascript/rev_comp.html

Forward Primer

5’ ATGCTTAACAGAACAATTGC 3’

Length = 20

Tm = 46°C

GC% = 35%

Reverse Primer

5’ TTACATGGTAGATAATTTGTTG 3’

Length = 22

Tm = 46°C

GC% = 27%

IDH2

Forward Primer

5’ ATGTTGAGAAATACTTTTTTTAG 3’

Length = 23

Tm = 45°C

GC% = 22%

Reverse Primer

5’ TTATAATCTCTTGATGACTG 3’

Length = 20

Tm = 44°C

GC% = 30%

IDP1

5’ ATGAGTATGTTATCTAGAAG 3’

Length = 20

Tm = 44°C

GC% = 30%

5’ TTACTCGATCGACTTGATTT 3’

Length = 20

Tm = 46°C

GC = 35%

Forward Primer

5’ ATGACAAAGATTAAGGTAGC 3’

Length = 20

Tm = 46°C

GC% = 35%

5’ TTACAATGCAGCTGCCTCGA 3’

Length = 20

Tm = 52°C

GC = 50%

Forward Primer

5’ ATGAGTAAAATTAAAGTTGTTCAT 3’

Length = 24

Tm = 45°C

GC% = 21%

5’ TTATAGTTTGCACATACCTT 3’

Length = 20

Tm = 44°C

GC% = 30%

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