for a 5:1 insert: vector ligation reaction, including the volumes of insert and vector you calculated above, and the volumes required for a 20 uL reaction: L
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Note that the table provided shows a ligation using a molar ratio of 1:3 vector to insert. Write out the complete recipe for a 5:1 insert: vector ligation reaction, including the volumes of insert and vector you calculated above, and the volumes required for a 20 uL reaction:
- Ligase buffer
- Nuclease-free water
- T4 DNA ligase
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- what type of gel(in terms of material) can be more suitable for the electrophoresis of 500-1000bp DNA fragment and why?How much 340bp insert DNA should be added to a ligation in which 50ng of 4.3kb vector will be used? The desired vector: insert molar ratio needs to be 1:3.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
- Could you please explain these: 1. Why analyze the unknowns in a (short) serial dilution rather than at a single concentration? 2. Which technique can detect lower concentrations of DNA? Compare the detection limit of the fluorescence approach and the absorbance approachIn bacterial transformation ... 1, Why do you have to incubate the competent bacterial cells on ice after the addition of plasmid DNA? 2. Why do you have to add SOC (Super Optimal Broth with Catabolite repression) medium to bacterial cells? 3. Why do you have to incubate the bacterial cells in SOC medium at 37oC for 45 minutes?That's the result of Gel electrophoresis of genomic DNA ( Of genomic DNA extraction experiment), please discuss the results and label and name the image to illustrate the answer? - Marker band sizes in gel: From top (well side) to bottom the bands have the following size in base-pair/bp- 6751,3652,2827,1568,1118,825,630
- Questions are talking about amplification of DNA When we run the gel, we also run a ladder why? what information does that give us. why does it form a ladder shape on the gel (what is happening to cause things to separate out in many bands)?1d) 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?Draw a gel to represent the band shift assay result for testing the following mixtures: (1) Eukaryotic DNA along(2) Eukaryotic DNA + TFs(3) Eukaryotic DNA + RNA polymerase II(4) Eukaryotic DNA + TFs + RNA polymerase II(5) Eukaryotic DNA + TFs + RNA polymerase II + nucleotidesBands for smaller molecules can be ignored
- Approximately how many high-energy bonds does DNA polymerase use to replicate a bacterial chromosome (ignoring helicase and other enzymes associated with the replication fork)? compared with its own dry weight of 10–12 g, how much glucose does a single bacterium need to provide enough energy to copy its DNA once?How would I solve this? primer (5’ TCAAAACG 3’ ) is shown hybridized to its template DNA below. Let’s say this DNA is used in a Sanger Sequencing reaction. How long (in nt) will dev the shortest RED nucleic acid chain be given that the red fluorophore is attached re to the ddCTP?Can you please help with 1f please picture with 1 graph is for question 1a) picture with 4 graphs is for question 1b) 1a) E. coli DNA and binturong DNA are both 50% G-C. If you randomly shear E. coli DNA into 1000 bp fragments and put it through density gradient equilibrium centrifugation, you will find that all the DNA bands at the same place in the gradient, and if you graph the distribution of DNA fragments in the gradient you will get a single peak (see below). If you perform the same experiment with binturong DNA, you will find that a small fraction of the DNA fragments band separately in the gradient (at a different density) and give rise to a small "satellite" peak on a graph of the distribution of DNA fragments in the gradient (see below). Why do these two DNA samples give different results, when they're both 50% G-C? 1b) If you denatured the random 1000 bp fragments of binturong DNA that you produced in question 1a by heating them to 95ºC, and then cooled them down to 60ºC and…