* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Example Quiz
DNA sequencing wikipedia , lookup
Mitochondrial DNA wikipedia , lookup
Zinc finger nuclease wikipedia , lookup
Nutriepigenomics wikipedia , lookup
Comparative genomic hybridization wikipedia , lookup
Microevolution wikipedia , lookup
Site-specific recombinase technology wikipedia , lookup
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing wikipedia , lookup
Primary transcript wikipedia , lookup
DNA profiling wikipedia , lookup
Cancer epigenetics wikipedia , lookup
Point mutation wikipedia , lookup
DNA polymerase wikipedia , lookup
Bisulfite sequencing wikipedia , lookup
SNP genotyping wikipedia , lookup
Genome editing wikipedia , lookup
Vectors in gene therapy wikipedia , lookup
DNA damage theory of aging wikipedia , lookup
Genealogical DNA test wikipedia , lookup
Gel electrophoresis of nucleic acids wikipedia , lookup
United Kingdom National DNA Database wikipedia , lookup
Cell-free fetal DNA wikipedia , lookup
Non-coding DNA wikipedia , lookup
Therapeutic gene modulation wikipedia , lookup
Genomic library wikipedia , lookup
DNA vaccination wikipedia , lookup
Extrachromosomal DNA wikipedia , lookup
Epigenomics wikipedia , lookup
Molecular cloning wikipedia , lookup
Artificial gene synthesis wikipedia , lookup
History of genetic engineering wikipedia , lookup
Helitron (biology) wikipedia , lookup
Nucleic acid analogue wikipedia , lookup
DNA supercoil wikipedia , lookup
Cre-Lox recombination wikipedia , lookup
BIO 510 Quiz 1 WITH ANSWERS 1. (2 pts) DNA ligase differs from EcoRI in that its enzymatic reaction requires (circle all correct responses) a. Magnesium (in both) b. ATP (ligase only) c. Lead acetate (in neither) d. DNA substrate (required by both) e. Ethanol (in neither) 2. (2 pts) If the nucleotide composition of a DNA molecule is random (that is, no bias toward or against any of the 4 bases), on average how frequently would each of the restriction enzymes cut (for instance, 1/256, that is, once every 256 base pairs)? Only the top strand of the restriction site is given, in parenthesis is the enzymes name. a. 5’ GAATTC 3’ (EcoRI) 46 = 1/4096 b. 5’ GATC 3’ (Sau3A) 44 = 1/256 c. 5’GTYRAC 3’ (HincII) 44 X 22 = 1/1024 d. GACGCNNNNN (HgaI) 45 X 15 1/1024 Note: Y = C or T; R=A or G; N=A,C,G, or T 3. (6 pts total) After you digested the pTZ18u(or19u) vector with EcoRI and HindIII you added a mixture of phenol/chloroform/isoamyl alcohol (PCI) mixture to your digestion. a. (2 pts) Tell why you did this (what was the value to your experiment in doing this step)? The goal was to remove the restriction enzyme from the DNA mixture. This was important as the next step was to ligate this DNA with the insert. If the EcoRI or HindIII was still present it would compete with the ligase activity (i.e., ligase would join the ends and then the EcoRI would re-cleave the DNA). b. (2 pts) What biochemical principles underlie the basis for this technique (that is, why does it work)? This is a two phase partition between an aqueous layer and an organic layer. The hydrophilic nature of DNA makes it partition in the hydrophilic layer while the protein is found in the organic layer or at the interface between the organic and aqueous layers. c. (2 pts) What role did each of the organic compounds contribute to the PCI function? i. Phenol – denatures and solubilizes proteins ii. Chloroform – denatures protein iii. Isoamyl alcohol – helps create a distinct interface between the aqueous and organic layers