Team:Wash U/Protocol
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:Colony PCR is similar to regular PCR in that it is used to amplify DNA. We have used colony PCR to verify successful transformations by adding grown up cells into the PCR mix, lysing the cells, and then amplifying the DNA. The amplified DNA can then be run through a gel and the number of base pairs can then be compared to an expected result. | :Colony PCR is similar to regular PCR in that it is used to amplify DNA. We have used colony PCR to verify successful transformations by adding grown up cells into the PCR mix, lysing the cells, and then amplifying the DNA. The amplified DNA can then be run through a gel and the number of base pairs can then be compared to an expected result. | ||
'''Materials''' | '''Materials''' | ||
- | * AccuTaq LA DNA Polymerase | + | * 0.5uL AccuTaq LA DNA Polymerase |
* 5uL 10x Buffer | * 5uL 10x Buffer | ||
* 2.5uL dNTP mix | * 2.5uL dNTP mix | ||
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# Add 700uL of Wash Solution to the column and cetrifuge for 1 minute and discard the flow through. Centrifuge the column again for 1 minute to remove excess ethanol. | # Add 700uL of Wash Solution to the column and cetrifuge for 1 minute and discard the flow through. Centrifuge the column again for 1 minute to remove excess ethanol. | ||
# Transfer the column to a new collection tube. Add 50uL of preheated Elution solution (65C) to the column and centrifuge for 1 minute. The desired DNA is now in the collection tube. | # Transfer the column to a new collection tube. Add 50uL of preheated Elution solution (65C) to the column and centrifuge for 1 minute. The desired DNA is now in the collection tube. | ||
+ | [https://2009.igem.org/Team:Wash_U/Protocol Back To Top] | ||
+ | |||
<font size="4"> | <font size="4"> | ||
=='''Making Competent Cells'''== | =='''Making Competent Cells'''== | ||
<font size="2"> | <font size="2"> | ||
- | + | :description | |
+ | '''Materials''' | ||
+ | *bulleted list | ||
+ | '''Procedures''' | ||
+ | #numbered steps | ||
+ | [https://2009.igem.org/Team:Wash_U/Protocol Back To Top] | ||
Revision as of 22:41, 16 July 2009