roblem #3 (25 points). You run a biochemical analysis lab you have isolated a nu

Question

roblem #3 (25 points). You run a biochemical analysis lab you have isolated a nucleotide piece of rom an E. coli sample you were provided. You have determined that 76 percent this RNA is part of a coding of region, while the remaining percentage is the very end of a gene product and hence is non-coding An HPLC analysis of an the RNase-treated aliquot of the mRNA reveals that it contains about equal fractions G,C,A, and U. What is your best guess for molecular mass of this mRNA? Provide a reasonable extinction coefficient for this RNA for UV absorption at 260 nm and use it to calculate the micromolar concentration of your original sample if a 1/1000 dilution gives an OD,60 of 0.0126 in an Eppendorf Biophonomer with a 1 cm cuvette. Without any further analysis, how much information content (in bits) exists in the nucleotide sequence of the coding region? Similarly, how much information content (in bits) exists in the amino acid sequence generated from the coding region? Describe in as much detail as you can the methods you would use to determine the exact nucleotide sequence of this mRNA based on procedures described in class.

Explanation / Answer

mRNA length = 75 nucleotide.

now only 76 percent is coding means rest 24 % is poly A tail. now for ease of calculation

we assume that mRNA is 75 nucleotide and each have equal share. so 75/4 = 18 nucleotide.

which means 72 nucleotide and 3 are Adenine( or mix of three nucleotide). now molecular weight of this mRNA

76* 330 = 25080. (330 is the avg mw of a nucleotide).

now

A = Ecl, where c= concentration and l= path length, E= extinction coefficient and

nucleic acid conc. = OD260 * standard coefficient * dilution

conc = 0.0126 * 40ug * 1/1000 = 0.000504 ug.

now A = Ecl.or E = A/cl

E = 0.0126/ 0.000504 = 25.

now no of coding nucleotide are 75.

so 72 are coding and 3 are stop codon. means we got amino acid chain of 72/3 = 24 amino acid.

for exact nucleotide sequencing, you may perform nucleotide sequencing of mRNA.

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