Bio253L_Section1_Exam4_Fall2021

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Test 4 Biol 253L Fall 2021 Section 1 Name:___________________________________________________ Date:____________________________________________________ Tuyen Ta 12/10/21

There is a new RFLP locus identified for Brassica rapa named Park13. Below are the two possible alleles for this locus: SnpA and SnpD. Both alleles are the same total length (1470 bp), but they have different restriction enzyme cut sites when digested with SpeI. You will answer each question showing ALL work and draw the bands that would be expected if the DNA was run on a gel. (50 points total) Use the information below to determine the bands that would be seen for Lanes A - F. For each question below also answer: (1) how many fragment(s) of DNA would be expected, (2) each fragment length, and (3) the number of band(s) that would be seen on the gel. Remember the total number of fragments from the digest/PCR amplification might not be the same as seen on a gel. 247 311 229 351 432

Lane A: Draw the band(s) that you would see after fully digesting the PCR product of P1 that is homozygous for SnpA with SpeI. (10 points) Number of DNA fragment(s) expected: Number of band(s) observed: Length of each band: Lane B: Draw the band(s) that you would see after fully digesting the PCR product of P2 that is homozygous for SnpD with SpeI. (10 points) Number of fragment(s) expected: Number of band(s) observed: Length of each band: Lane C: Draw the band(s) you would see after fully digesting the PCR product of an offspring (F1) of P1 and P2 with SpeI. If there is more than one possible offspring genotype for the cross of P1 and P2, then select one possible genotype to draw on the gel. Show how you determined the possible genotypes of the F1 generation. (15 points) Possible F1 genotype(s): Number of fragment(s) expected: Number of band(s) observed: Length of each band: Lane D: Draw the band(s) you would see after running undigested PCR product of P1 that is homozygous for SnpA with SpeI. (5 points) Number of fragment(s) expected: Number of band(s) observed: Length of each band: 8 4 687 bp , 247 bp , 311 bp , 225 bp 6 3 687 bp , 351 bp , 432 bp D D AAD / AD " 00% AD genotype AAD AD y 6 687 bp , 247 bp , 311 bpi 225 bp , 351 bp , 432 bp 1 1 1470 bp

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Lane E: Draw the band(s) you would see after running undigested PCR product of P2 that is homozygous for SnpD with SpeI. (5 points) Number of fragment(s) expected: Number of band(s) observed: Length of each band: Lane F: Draw the band(s) you would see after running undigested PCR product of the offspring generation (F1) of P1 and P2 for the Park12 locus. (5 points) Number of fragment(s) expected: Number of band(s) observed: Length of each band: Exam total _____/ 50 i ' 1470 bp I 1 1470 lbp

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