Put the following pieces of DNA in order that they would appear reading from the top (nearest the loading point) to the bottom of a gel, with "first" meaning nearest the top. ( Choqgel First [Choose] 21,000 basepairs (bp) 10,000 bp 23 kilobases (kB) 18 kB Second 2,300 bp Choose ] Third [ Choose] Fourth [ Choose ] Fifth
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![Put the following pieces of DNA in order that they would appear reading from the top (nearest the loading
point) to the bottom of a gel, with "first" meaning nearest the top.
First
[ Choqse
[Choose]
21,000 basepairs (bp)
Second
10,000 bp
23 kilobases (kB)
18 kB
2,300 bp
Third
[Choose ]
Fourth
[ Choose]
[Choose]
Fifth
>
>
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- Suppose a piece of DNA is cut into four fragments of varying sizes. Where would you expect to find the larger fragments on the gel (top, middle, or bottom)? Why? Suppose you had 1000 pieces of each of the four fragments, how many bands would appear on the gel?You have 10 ul of DNA you want to cut with HindIII, and then run a gel, in order to cut out certain bands and purify the DNA from them. Since the DNA is pretty concentrated, you are going to cut it in a 50 ul total volume and then run it in 2 adjacent lanes of a gel, to avoid overloading the lanes. What are all the ingredients you need to put into a tube, how much of each, and what is the temperature ordinarily used? (Amount of HindIII might vary, but be safe and put in twice as much as you might normally use to cut less DNA)During agarose gel electrophoresis, why does DNA move through the gel when electric current is applied? because DNA is negatively charged because a charged chemical from the loading buffer is bound to the DNA because DNA is positively charged because DNA absorbs electricity
- The following are DNA fragments containing a small gene. The top strand is the coding strand. Transcribe all 5 groups and translate. Group A 5’-GGCAATGGGTTTGTGCAATTCTAAAAGTTTTTAATTC-3’ 3’-CCGTTACCCAAACACGTTAAGATTTTCAAAAATTAAG-5’ Group B 5’-GGCAATGGGTTTGTGAAATTCTAAAAGTTTTTAATTC-3’ 3’-CCGTTACCCAAACACTTTAAGATTTTCAAAAATTAAG-5’ Group C 5’-GGCAATGGGTTTGTGCAATTCTAAGAGTTTTTAATTC-3’ 3’-CCGTTACCCAAACACGTTAAGATTCTCAAAAATTAAG-5’ Group D 5’-GGCAATGGGTTTGTGCAATTCTAACAGTTTTTAATTC-3’ 3’-CCGTTACCCAAACACGTTAAGATTGTCAAAAATTAAG-5’ Group E 5’-GGCAATGGGTTTTGCAATTCTAAAAGTTTTTAATTC-3’ 3’-CCGTTACCCAAAACGTTAAGATTTTCAAAAATTAAGBased on your gel, how big are the DNA fragments in each lane? Enter your results in Table 2.When proteins are separated using native gel electrophoresis, size, shape, and charge control their rate of migration on the gel. Why does DNA separate based on size, and why do we not worry much about shape or charge?
- Total nucleic acids are extracted from a growing culture of yeast cells. They are then mixed with specialized beads to which the single-stranded DNA molecule with sequence 5’-TTTTTTTTTTTTTTTTTTTTTTTTT-3’ has been covalently attached to the surface (see image to the right, where each black line represents a polynucleotide sequence). After a short incubation time, the beads are removed from the mixture. When you analyze the cellular nucleic acids stuck to the beads, which type of nucleic acid (i.e. DNA, rRNA, etc.) do you expect to be the most abundant? Why?Given the following DNA sequence: 5’-ATGCGGCCAAGGTCAGAGTGACA-3’ a) If this DNA strand represents the “Sense Strand” of DNA, what would be the RNA sequence? b) If this DNA strand represents the “Antisense Strand” of DNA, what would be the RNA Sequence? c) What would be the other strand of DNA?A DNA fragment with 450 bp will be closer to the top (negative pole) of an electrophoresis gel than one with 2,500 bp.True or false?
- Whether done manually or automated, DNA sequencing gels are always made of polyacrylamide rather than agarose. Why can't agarose be used for a sequencing gel, as it is for other DNA gel electrophoresis?A DNA synthesizer “machine” is used to create short single stranded DNA of any given sequence. You have used the machine to create the following the DNA molecules: (DNA #1) 5’- CTACTACGGATCGGG – 3’ (DNA #2) 5’- CCAGTCCCGATCCGT – 3’ (DNA #3) 5’-AGTAGCCAGTGGGGAAAAACCCCACTGG-3’ Now you add the DNA molecules either singly or in combination to reaction tubes containing DNA polymerase, dATP, dCTP, dGTP, and dTTP in a buffered solution that allows DNA polymerase to function. For each of the reaction tubes, indicate whether DNA polymerase will synthesize any new DNA molecules, and if so, write the sequence(s) of any such DNAs. DNA #1 plus DNA #3 DNA #2 plus DNA #3 DNA #1 plus DNA #2 DNA #3 onlyWrite the complementary sequence for the following DNA sequence, in order from 3' to 5': 5′−CGATATTGAGCTAAGCTT−3′