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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- Draw the gel electrophoresis results for the following 16 kb plasmid when the plasmid is cut withrestriction enzyme A, with restriction enzyme B and with both A and B restriction enzymes. The Aenzyme cuts at positions 0 and 9.2 kb and the sites are shown as white. (0 is at the top of the circle.) TheB enzyme cuts at positions 3.8, 7 and 13.2 kb and the sites are shown as gray. Show the electricalpolarity of the gel, the bands, and their sizes in your drawing. Make sizes consistent between gel lanes.Explain why the concept of charge density is important for gel electrophoresis.Kha Vu Danels Include: 8DX : Safehy Jor bromie tnto lab repart! Name Section date sheet MAPPING PRACTICE #1 Below is a restriction map for the plasmid PGEN 101 (total length = 20 Kb). Using this map as a guide, give the number of restriction fraqments along with their associated lengths that would result from digesting PGEN 101 with the restriction enzymes EcoRI, BamHI and a combination of ECORI and BamHI. BamHI 3.2 Kb 1.7 Kb EcoRI BamHI PGEN 101 8.7 Kb 5.5 Kb .9 Kb EcoRI ECORI DIGESTION PERFORMED SIZES OF FRAGMENTS OBTAINED 10.4 kb , 0.9kb, 8.7 Kb EcoRI 3.2 Kb, 16. 8kb BamHI EcoRI + BamHIThe following are the restriction enzyme sequences. The slash indicates the cut in the DNA sequence. EcoRI: 5' G/AATTC 3' Hind IlI: 5' AG/GATCC 3' Bam Hl: 5' G/GATCC 3' For DNA sequences 1,2 and 3 (below) Write the 3' to 5' DNA bottom strand • Write the mRNA sequence of top strand Write the mRNA sequence of bottom strand Write the Protein sequence of the Bottom strand • Which enzyme should be used to differentiate (detect the mutation) in sequence 2 1) cac agt aca tca gac atg gat cca agc cca tgt ata ccc cg aac 2) cac agt aca tca gac atg gtT cca agc cca tgt ata ccc ccg aac 3) cac agt aca tca gac atg gaC cca agc cca tgt ata ccc ccg aac Edit View Insert Format Tools Table 12pt v Paragraph
- Restriction Enzymes Handout plasmid Restriction Enzymes Bam HI Eco RI Hpa I Hind III Nde I Please solve this question in less than 15 minutes for genetics engineering Sal I Plasmid Handout DNA Sequence (both strands are represented) GIGATCC CCTAG G U G|AATTC CTTAA G GTT | AAC CAAjTTG A|AGCTT TTCGA | A CATATG GTATAC G|TCGAC CAGCT | G agtgacata tga ttcgag c cggta a c tca ctgtat a ст aagctcgagccattg cggggat cctctag agtcgacctgcagg c gcccctaggagatctcagctgga cgtc cg tagca ag c tggcgt at atggta cata at cgttcga accgcattagtaccat gta t gggatceptet ct ccagtaggtaggc cgtcg ccct agga aga g g t catccat cc ggcag c Antibioty Resistance gene gc taggetta a actgggatccat gcc origin o tplgm cg at cc a at ttgaccctaggta cgplasmid replication What is the role of the plasmid ? Explain the experimental details of the transformation. How can the success of transformation be observed ?You are studying a new plasmid, and you digest the plasmid with three restriction enzymes: Eco RI (E), HindlII (H), and Xbal (X). You digest the plasmid DNA with each of the following combinations of enzymes and observe the results on an agarose gel. You are provided a partial plasmid map as shown below to the right. E H E+H E+X H+X Kb +4.3 +2.8 +2.5 - +2.0 -- -1.8 -1.5 12 +1.0 +0.8 +0.5 - a. What is the size of this plasmid in base pairs? b. What is the distance in base pairs between E1 and H? c. What is the distance in base pairs between E1 and X? d. What is the distance in base pairs between E2 and H? e. What is the distance in base pairs between E2 and X?Below is a plasmid with restriction sites for Baml and EcoRI, Several restriction digests were done using these two enzymes either alone or in combination. Hint: Begin by determining the number and size of the fragments produced with each enzyme. "kb" stands for kilobases, or thousands of base pairs. Plasmid Gel lanes I | III IV V 6 Kb Bam HI Bam HI. 20 Kb PGEN101 (20 Kb) 2 Kb 11 Kb Bam HI 8 Kb 8 Kb 4 Kb 6 Kb Eco RI 3 Kb 21. Which lane shows a digest with BamHl only? a. I b.I c. II d. IV e. V 22. Which lane shows a digest with EcoRLonly? a. I 23. Which lane shows the fragments produced when the plasmid was incubated with both EcoRI and BamH1? a. I b.I c. II d. IV e. V b. I c. II d. IV e. V Base pairs
- A piece of DNA fragment is sequenced. You clone the the fragment, isolate the cloned DNA fragment, and set up a series of four dideoxy reaction. You then separate the products of the reaction by gel electrophoresis and obtain the following banding patter: ddATP ddTTP O 5'-ATTCGACT-3' O 5'-TCAGCTTA-3' What is the base sequence of the synthesized fragment? O 5'-AGTCGAAT-3' ddCTP O 5'-TAAGCTGA-3' I ddGTPetion enzyme Sall cuts the sequence GTCGAC to leave a five base, 5' overhang. The restriction enzyme Xhol The he sequence CTCGAG to leave a five base, 5' overhang. You mix a Sall-digested insert with a Xhol digested plasmid vector and perform a ligation. Draw your Sall digested insert and your Xhol digested plasmid vecior. Then ligate them in place and draw the insert in the vector. Use two different colors to distinguish the vector and the insert. Does your insert ligate into the vector in a specific orientation? Yes, or no? Explain.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?
- U have the plasmid pUC18/19, which is a circular plasmid that consists of 2686 bp. What would the number of and length of the fragments be if you cut the plasmid with the following restriction enzymes or combination of enzymes? Give a schematic representation of the digestions. PscI & GsuI ______________________________________________________________ ScaI, PdmI & BsaXI ______________________________________________________________ ScaI, SspI & EheI ______________________________________________________________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 DNAA 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.