User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/07/24: Difference between revisions
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**# Extremely unspecific amplification. Size indeterminable. | **# Extremely unspecific amplification. Size indeterminable. | ||
***CONCLUSION: Longer N-mer primers with higher annealing temperatures lead to higher but more unspecific amplification. It looks as if the size of the N-mer needs to be optimized with specificity, and something like a 34-mer may be the best choice. However, I note in this experiment, I used cloned PFU DNAP, not high fidelity PFU DNAP. I will need to repeat this experiment with HF PFU DNAP. In the meantime, I can still optimize primer concentration using the 45-mer. | ***CONCLUSION: Longer N-mer primers with higher annealing temperatures lead to higher but more unspecific amplification. It looks as if the size of the N-mer needs to be optimized with specificity, and something like a 34-mer may be the best choice. However, I note in this experiment, I used cloned PFU DNAP, not high fidelity PFU DNAP. I will need to repeat this experiment with HF PFU DNAP. In the meantime, I can still optimize primer concentration using the 45-mer. | ||
* Performed [http://openwetware.org/wiki/Corum:Whole_Plasmid_PCR whole plasmid PCR] with ΦX174 primer 1 (45-mer) with range of primers: 0, 100, 200, 1000, 2000 nM. | |||
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Revision as of 16:34, 24 July 2012
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Hypothesis 2: Gene L is necessary for phage propagation.
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