Pre-lab (Week 12)

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School

California State University, Fullerton *

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Course

253L

Subject

Biology

Date

Dec 6, 2023

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pdf

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1

Uploaded by MinisterSnowRhinoceros34

Pre-lab questions for Biol 253L, week 12. The answers are found in this week’s reading. Answer in the space below and continue on back if necessary. 1. What is the definition of “Microsatellite DNA”? Microsatellite DNA refers to the region of DNA that has a pattern of short tandem sequences. 2. What does the acronym “VNTR” stand for? VNTR means variable number tandem repeats. 3. How are the VNTRs commonly used to study B. rapa different from microsatellite DNA? VNTRs are commonly used to study B. Rapa because they help differentiate one plant from another, similar to how humans have unique fingerprints that set us apart from others. This is different from micro satellite DNA because micro satellite is a type of VNTR while VNTR can be it’s own thing. 4. What is the name of the locus that you are PCR amplifying this week? On which chromosome is it found? How do the two possible alleles at this locus differ from each other? Based on the lab manual, the PCR we will be amplifying in lab this week is the D9 B.rapa S1 which is located in chromosome 9. However, it’s stated that even though its’s not a VNTR, the two possible alleles don’t have a different number of sizes because of a VNTR. But, one of the alleles has a 53-bp insertion right after the tandem will repeat. 5. What is in the “PCR Master mix” that you are using for your PCR reactions? The PCR master mix included dNTPs, Tap polymerase buffer, Tap polymerase, and the DNA primers ; D9 BrapaS1 Forwards + Reverse, and the loading dye. 6. How many PCR tubes will you be using this week? How will each tube differ from each other? According to the lab manual, we should label one PCR tube for all of the three differing DNA template samples. Each tube will differ from each other based on the DNA sample inside. 7. List the specific temperatures are you using for the denaturation, annealing, and extension steps for your PCR. Denaturation: 95 C for 30 seconds Annealing: cooled at 50 C for 30 seconds Extension: heated to 72 C for 30 seconds for every 500 bp
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