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From 2010.igem.org

Ligation of Reporter and Terminator (About two hours)

We used the same quantities that were used on September 23 2010

PCR (About 2.5 hours)

On October 1, we did the miniprep of the cloning product of the gene Rh1ab in the pGEM-T vector

Today, we will do a PCR of this to see whether the clones that have grown contain the gene Rh1ab

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No bands appear

Possible reasons:

1) The PCR has had some sort of problem

2) The cloning has had some sort of problem

Possible solutions:

1) Repeat the PCR, changing the denaturizing temperature time to 8 minutes and in the cycling from 30 seconds to 1 minute. Besides that, we used two different miniprep samples from the cloning (maybe the colony that we picked doesn't have the insert)

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