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Team:Tsinghua/Notebook/31 July 2010
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(Difference between revisions)
(New page: == Module I, Group 2c == To solve the problem of severe dimerization of primers, we tried to adjust PCR system and use different concentration of upstream and downstream primers. As the c...) |
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PCR system (pyrobest): | PCR system (pyrobest): | ||
H2O 17.3ul | H2O 17.3ul | ||
| - | + | 10×buffer 2.5μl | |
dNTP 2.5μl | dNTP 2.5μl | ||
primer1 1μl | primer1 1μl | ||
primer2 0.5μl | primer2 0.5μl | ||
template 1μl | template 1μl | ||
| - | + | pyrobest 0.2μl | |
| - | Total | + | Total 25μl |
---- | ---- | ||
Result: to my disappoint, after gel purification,target fragments disappeared! What's worse, no PCR product of Chlr is seen in the gel photograph actually. | Result: to my disappoint, after gel purification,target fragments disappeared! What's worse, no PCR product of Chlr is seen in the gel photograph actually. | ||
Latest revision as of 16:58, 26 October 2010
Module I, Group 2c
To solve the problem of severe dimerization of primers, we tried to adjust PCR system and use different concentration of upstream and downstream primers. As the concentration of PCR product is too low to be digested and purification, we attempted to use PCR product got yesterday as the template for PCR today.
PCR system (pyrobest):
H2O 17.3ul 10×buffer 2.5μl dNTP 2.5μl primer1 1μl primer2 0.5μl template 1μl pyrobest 0.2μl Total 25μl
Result: to my disappoint, after gel purification,target fragments disappeared! What's worse, no PCR product of Chlr is seen in the gel photograph actually.
