1. If a restriction enzyme cuts a circular plasmid twice, how many fragments would you see on the gel?
2. How would you estimate the total number of base pairs in a plasmid by looking at the DNA fragments of the digested plasmid on a gel?
3. If a linear 1kb DNA fragment has a restriction site that is located 50 bp from one end of the plasmid, what would you expect to see if the digested and undigested DNA samples were run on a 1% agarose TAE gel?
1)
If a restriction enzyme cut a plasmid two times,then two fragments are obtained.
Look in the image for diagrammaric representation:
2)
We also load a DNA ladder in the well of gel electrophoresis. We also load our digested sample in the gel. Then look for the corresponding band fragments. And compare it with the DNA ladder. In this way, the molecular weight of fragments are obtained.
3) There is found one restriction site in the plasmid. That's why the digested plasmid have linear DNA.
The uncut plasmid have two forms open circular DNA and supercoiled DNA. The following pattern is observed in the gel.
Supercoiled DNA runs faster than open curcular DNA because of less friction.
Please rate.
1. If a restriction enzyme cuts a circular plasmid twice, how many fragments would you see...
1) If a restriction enzyme cuts a circular DNA into five fragments, how many restriction sites are there in the DNA? 2) How many molecules of DNA will be present after 6 cycles of PCR, if you started with one double-stranded DNA molecule? CELL BIOLOGY QUESTIONS!! SHOW WORK PLEASE
A restriction digest of a circular plasmid is conducted using two different restriction enzymes. Restriction enzyme 1 cuts the plasmid in one place while restriction enzyme 2 cuts in two places. All restriction sites are at least 1kB apart from each other. Based on this information, the resulting gel should have _____ bands for the enzyme 1 reaction, _____ bands for the enzyme 2 reaction, and _____ bands for the reaction that contains both enzymes A 1; 2; 3 B...
In your previous prac session you digested your POTC-A plasmid DNA with 3 enzyme mixes (AB and C). One mix contained the restriction endonuclease Kpnl alone, another contained both del and Not and another contained Sphi and Nhe, but you don't know which was which yet. The sites where these enzymes cut POTC-A are indicated on the plasmid map at the top of the page. Calculate the sizes of the DNA fragments that you would expect to see on your...
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers to locations where the restriction endonuclease EcoRI cuts the DNA, and Pst refers to locations where the restriction enzyme Pst cuts the DNA. Potential restriction sites are numbered 1-6. Distances between restriction sites are shown on the bottom scale in base pairs (bp). The thick line represents the part of the molecule that has homology with a probe. Eco Pst Eco Pst Eco Pst...
Now determine the sizes of the fragments generated after digesting the r-plasmid with each of the restriction enzymes or combination of restriction enzymes. r-plasmid digested with Pstl r-plasmid digested with EcoRI r-plasmid digested with both Pstl and EcoRI Question 8. (3 pts.) Use this information to determine the relative placement of these restriction sites in the r-plasmid, and then draw restriction site maps of the vector and the r-plasmid on the following circular chromosome templates. The map should show the...
can someone explain throughly on how to find a-c??? thanks!!! The following question will provide practice in interpreting and analyzing gel results. 5. You obtained the DNA electrophoresis gel below. Three samples of lambda phage DNA were digested with 3 different restriction enzymes and the digested DNA was applied to the gel in lane 4 and the bands were visualized. The Hind Ill digest was used as a molecular weight standard marker and produced 6 DNA fragments of known size:...
Prac result Nhel Kpn I 1569 1601 Kpni 1030 2295 / 2310 Not I 988 ОТС-д Sphi 2559 Nde I 484 POTC-A 6401 bp Hygro Amp Nde1 3369 pUC Ori 5260 4587 Amp: Ampicillin resistance gene 5405 - 6265 Hygro: Hygromycin resistance gene Choose the gel image with correct bands in lanes 1 to 5 to indicate the approximate positions where you expect to see these predicted DNA fragments. (1 mark) Lane 1: POTC (undigested / uncut) Lane 2: POTC-A...
Suppose you are going to do a restriction digest with a plasmid, using the restriction enzyme Eco R1. A map of the plasmid is shown here. The entire plasmid is 6000 bp, and there are Eco R1 restriction sites at 1500 bp, 2000 bp, and 4000 bp. You’re going to run the entire volume of the digest on a gel, and you want to cut just enough DNA to have 50 ng in the smallest band on your gel. Starting...
Hi I have a problem with number 5, it involves gel analysis results. There are 2 parts, a,b,c. For A Im sure you need to make a graph with distance in (cm) on the vertical axis and log10 bp on the horitzontal. I need help figuring out where to start and what to do. Please help! The following question will provide practice in interpreting and analyzing gel results You obtained the DNA electrophoresis gel below. Three samples of lambda phage...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...