What is the dna strand sequence for phosphate sugar backbone?
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What is the dna strand sequence for phosphate sugar backbone?
What are sticky ends in restriction enzymes
can you examples to help me understand.
Does where you cleave affect the length of the sticky ends overhang?
5-3directionalytgg atc gat atc gcc aat.
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- Cloning Genes Is a Multistep Process The following DNA sequence contains a six-base sequence that is a recognition and cutting site for a restriction enzyme. What is this sequence? Which enzyme will cut this sequence? (See Figure 13.5 for help.) 5 CCGAGGAAGCTTAC 3 3 GGCTCCTTCGAATG 5Describe the nature of recognition sites for restriction enzymes and the nature of the ends of the DNA that are left. Why do we need to run a gel electrophoresis after enzyme digestionOn the gel shown below are four DNA samples. Samples A to C are taken from tissues of landslide victims that are being identified, while sample D came from a hair sample brought by a mother looking for the remains of her son. (see img) i. If similar band patterns in a gel are created using the same restriction enzyme, what does that tell you about the DNA sequence of the samples? ii. In sample C, only two fragments were created. How many restriction sites (regions where enzymes cut) are present in sample C?
- Restriction enzymes and DNA ligase play essential roles in DNA cloning. How is it that a bacterium that produces a restriction enzyme does not cut its own DNA? Describe some general features of restriction sites.The partial sequence of one strand of a double-stranded DNA molecule is5′ – – – GACGAAGTGCTGCAGAAAGTCCGCGTTATAGGCATGAATTCCTGAGG – – – 3′The cleavage sites for the restriction enzymes EcoRI and PstI are shown below.Write the sequence of both strands of the DNA fragment created when this DNA is cleaved with both EcoRI and PstI. The top strand of your duplex DNA fragment should be derived from the strand sequence given aboveAn E. coli genomic DNA fragment is run out on a gel undigested (U), and then with two different restriction enzymes. Based on the gel below, what is the most appropriate reason for the and pattern displayed? DpnI is likely a restriction enzyme naturally found in E. coli. The DNA fragment is methylated at the EcoRI cut site, but not the DpnI cut site. EcoRI targets methylated cut sites. There is no DpnI cut site on the DNA fragment. The DNA is methylated at the DpnI cut site, but not the EcoRI cut site.
- Which of the enzymes from the following list wouldyou need to make a recombinant DNA molecule? Whatis the function of those enzyme(s) in the process?a. DNA polymeraseb. RNA polymerasec. A restriction enzymed. DNA ligasee. An aminoacyl-tRNA synthetasef. Peptidyl transferaseg. Reverse transcriptaseA student wanted to load .75 ug dna on agarose gel and has 4x loading buffer for sample preparation. The student has 50ul purified plasmid, finding concentration of plasmid to be 250 ng/uL The student used 10uL plasmid DNA in 50uL reaction for the restriction digest Give the volumes of restriction digest and 4x loading buffer that student would mix together and load on agarose gel.What sequence do the restriction enzymes used in the lab recognize? How do they cut? And how would these different cuts effect cloning? (i.e. compare and contrast how overhang and blunt cuts will differ with cloning.) What organisms were the restriction enzymes used in this lab derived from? Why are restriction enzymes important for the organism that makes them? What is the purpose of multiple cloning sites on pUC19? What size were each of your plasmid fragments?
- You digest 4 uL of plasmid DNA that is 50 ng/uL concetration in a total volume of 20 uL. You run 10 uL of the digest on teh gel. You then do a DNA purification protocol with a Zippy prep on the remaining digested DNA. You elute the DNA in a 25 uL. A 2 uL ssample has the concentration of 2 ng/uL. What is the DNA yield? YIELD = How much dna came out of the column/how much DNA was loaded onto the columnRemember to subtract amount of DNA run on gel from total digested DNA to get hw much DNA was loaded onto the columnAmount that came out of the column equals the volume of the eluted DNA times the concentration of the purified DNAYou digest 4 uL of plasmid DNA that is 50 ng/uL concetration in a total volume of 20 uL. You run 10 uL of the digest on teh gel. You then do a DNA purification protocol with a Zippy prep on the remaining digested DNA. You elute the DNA in a 25 uL. A 2 uL ssample has the concentration of 2 ng/uL. What is the DNA yield?EcoR1 is a restriction enzyme that cuts DNA at the following base sequence: 5' GAATTC 3' Which statement is true? EcoR1 must bind to the exon region of a gene. EcoR1 must bind to the coding strand of the DNA. This restriction site is not a palindrome sequence. EcoR1 cuts DNA but not RNA.