4. You will estimate the quantity of DNA present in the pUC18 DNA prep by gel ba

Question

4. You will estimate the quantity of DNA present in the pUC18 DNA prep by gel band brightness. Compare your linear plasmid band in the HindIII digest lane to the -HindIII marker. If, for example, your linearized plasmid is half as bright as the 4, 361 bp band of the marker, then your approximate DNA quantity is 90/2 or 45 ng. You loaded 10 l of the HindIII digest onto the gel so your original concentration of DNA in the digest is 45 ng/10 l or 4.5 ng/l. You put 10 l of DNA in your digest. All together then, you have 4.5 ng/l x 10 l or 45 ng of DNA in the digest.

I am comparing my linear Hind III digest plasmid to Fragment ____ of the -HindIII digest. 46

The amount of DNA in the linearized plasmid is ________ and I reach this number because_________________________________________________________________.

The original concentration (g/l) of pUC18 plasmid DNA in the HindIII digest is __________________.

The total amount of pUC18 plasmid DNA in the HindIII digest is ______________g.

The pBR322-BstNI digest cannot be used for quantitation but can be used to estimate fragment size. You may find this marker useful in analyzing your data (but quantitate the DNA using the lambda-HindIII digest marker).

Tube I: no enzyme

Tube II: 1 l HindIII

Tube III: 1 l HindIII, 1 l ScaI

Tube IV: 1 l BglI, 1 l ScaI

Explanation / Answer

The photo that you have provided shows the result of different digestions than those mentioned in the theory part of your questions or more image is required. Please check the image and get back to me. I will try to help you then.

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