Matt Gethers/CRI, Thailand/Labwork/Isolating HmgR/Week of 6.8.08
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6.11.08
Primer Design for HmgR ORF
Here is the KEGG File for HmgR (pa2010). I'd like to amplify this ORF and then drop it into the pET-11a commercial vector between the NdeI and BamHI sites. The ORF has to be in frame and I have to be sure there aren't any cloning sites in the ORF.
Using the file of HmgR from Mayuree
Forward Primer BT2736 (5' to 3')
| Extra Bases (For cutting efficiency) | NdeI Site | Annealing Bases |
| gcg | catatg | gaaaagaacagcagtccc |
Specs
LENGTH (annealing/all): 21/27
GC CONTENT (not including non-anealing bases):47.6 %
MELT TEMP (not including non-anealing bases): 55.4 ºC
Min(ΔG): -0.07 kcal.mole-1
Reverse Primer BT2737 (5' to 3')
| Extra Bases (For cutting efficiency) | BamHI Site | Annealing Bases |
| cgc | ggatcc | gctcggagactgcgtcat |
Specs
LENGTH (annealing/all): 18/27
GC CONTENT (not including non-anealing bases): 61.1 %
MELT TEMP (not including non-annealing bases): 57.4 ºC
Min(ΔG): -4.01 kcal.mole-1
