Concept explainers
To analyze:
Why the positions of
Introduction:
Blotting is the technique routinely used for the detection of
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Genetic Analysis: An Integrated Approach (2nd Edition)
- Describe what each of the 12 bands on your Western blot should have been. Remember, the 12 bands will be 4 conditions x 3 proteins (phospho-S6, phospho-AMPK, tubulin). Please describe the relative density of each band compared to the control (for example, how dense will phospho-S6 be in each of the three experimental conditions compared to the control condition?). For each band, provide a well-reasoned rationale for your anticipated result. Give the reason why cell signaling would produce each result in each condition.arrow_forwardii only.........arrow_forwardPlease explain all parts to questionarrow_forward
- Southern and Northern blotting are powerful molecular biological tools based on hybridization of nucleic acids. How are these techniques the same? How do they differ? Give some specific applications for each blotting technique.arrow_forwardProvide a detailed description and hand drawn figure of each of the following. (1) Mismatch Repair (2) Nucleotide-Excision Repairarrow_forwardantibodies were produced in a rabbit against a 25 KDa human soluble protien. when the rabbit antiserum was used in a western blot of human soluble protiens, it detected 25-KDa protien,but also bound to protiens of 65KDa and 39 KDa.assuming that a pure protien was used to make the antibodies,how do yuo explain the results of the western blot?arrow_forward
- In Colony Polymerase Chain Rxn (PCR), what primers are necessary to identify that the transformants contain the gene of interest with the correct orientation?arrow_forwardAnalysis of a 49kDA protein is aimed with a western blot technique. For this purpose, "whole cell extract" was obtained from biopsy samples taken from three different individuals, and when their spectrophotometric measurements were made, their concentrations were determined to be 40 µg/µL, 50 µg/µL, and 100 µg/µL for three samples, respectively. At the beginning of the experiment, 200 µg of protein is loaded into each gel well by using a ready 5x loading solution containing SDS and β-mercapta-ethanol together with glycerol and methylene blue dye. a) If loading will be done in a volume of 20µL, then how is the preparation of the samples before loading?b) Considering the loading content given above, what can be said about whether this PAGE is "non-denaturing" or "denaturing". Justify taking into account the loading solution content.arrow_forwardDiagram and explain how APEX probes can be used to determine that an individual is CC (homozygous) for a specific G/C SNV. Recall that the genotype of an SNV is identified from the strand shown in NCBI. What color fluorescence will be observed? Also, explain why a left apex probe cannot be used for this SNV. The SNV sequence, on the strand shown in NCBI, and a few nucleotides adjacent to the SNV are below: 5'-------TGT(G/C)CAG------3'arrow_forward
- You run a Western blot to detect Ras in sampe A and sample B. You find that sample B gives a darker band. What is the most appropriate interpretation of this? (a) sample A has more Ras DNA, (b) sample B has more Ras DNA, (c) sample A has more Ras mRNA, (d) sample B has more Ras mRNA, (e) sample A has more Ras protein, (f) sample B has more Ras protein. I understand that those molecules have higher molecular weight should give darker band. As mRNA has higher molecular weight than that of DNA and protein, I wish to pick (d) "sample B has more Ras mRNA" as the best answer. I am not sure if I am right. Please the expert advise.arrow_forwardWhy are there multiple results returned when you input a sequence into BLAT or BLAST? How do you know which is most relevant? Does that make the others irrelevant?arrow_forwardIn this western blot, the levels of phosphorylated TBK (PTBK) decrease with increasing amounts/expression of the viral protein (VP). Figure description: Increasing amounts of a plasmid expressing the viral protein (0.5, 1, or 2ug) were cotransfected with TBK1 expression plasmid. Cells were harvested 24 h post-transfection and analyzed for phosphorylated TBK1 (anti- TBK1 Ser172), total TBK1 (anti-TBK1), B-actin (anti-B-actin), and viral protein (anti-VP) expression by Western blot analysis. VP PTBK1 (S172) TBK1 actin Virus proteins O True O Falsearrow_forward
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