You will estimate the quantity of DNA present in the pUC18 DNA prep by gel band

Question

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 µl of the HindIII digest onto the gel so your original concentration of DNA in the digest is 45 ng/10 µl or 4.5 ng/µl. You put 10 µl of DNA in your digest. All together then, you have 4.5 ng/µl x 10 µl or 45 ng of DNA in the digest.

I am comparing my linear Hind III digest plasmid to Fragment ____ of the -HindIII digest.
The amount of DNA in the linearized plasmid is ________ and I reach this number because_________________________________________________________________. The original concentration (µg/µl) of pUC18 plasmid DNA in the HindIII digest is __________________. The total amount of pUC18 plasmid DNA in the HindIII digest is ______________µg. 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).
10,000 bp 3,000 bp 2,600 bo 1,000 bp 700 bo 250 bp

Explanation / Answer

I am comparing my linear Hind III digest plasmid to Fragment 4361bp band (fragment 4) of the -HindIII digest. The amount of DNA in the linearized plasmid is 45 ng and I reach this number because the assume band looks about half as bright as compared to fragment 4 of the -HindIII digest. The original concentration (µg/µl) of pUC18 plasmid DNA in the HindIII digest is 4.5 ng/l or 0.0045 µg/µl . The total amount of pUC18 plasmid DNA in the HindIII digest is 45 ng (4.5 ng/l x 10l = 45ng) or 0.045 µg. 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).

Related Questions

Navigate

• Browse (All)
• Browse Y
• Subjects

---

---

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.