Mark Nicholas L.

Yow

3B-BC

1. How many possible primer pairs were generated by Primer-BLAST? List the top 3 primer pair
sequence with long possible PCR product.
- There are 50 possible primers that can be generated by the Primer-BLAST.

Primer 1:

Forward primer – AAGGGGACTGGGGCTTCTAT Reverse primer – AACCATACACACCTCGCTGG

Primer 2:

Forward primer – CTGAATAGGTGACCGGAGGC Reverse primer – CAGGCGAGTTGTAGACCCAG

Primer 3:

Forward primer - GCCTGAATAGGTGACCGGAG Reverse primer – GGCGAGTTGTAGACCCAGTC

2. What is the average primer length generated? What is the highest and what is the lowest?
Answer in base pair (bp).
- All primers generated 20 base pairs which makes the average length 20 base pairs and the
highest and lowest length of primer is also 20 base pairs.

3. What is the most common GC content %? What is the highest and what is the lowest?
- The most common GC content % is 55% which has 11 results. The highest GC content % is
60% and the lowest GC content is 50%.
4. What is the range of Tm among the primers generated? Choose primers with Tm difference of
<5. List the top 3.
- The range of Tm among the primers generated is from 59.48 to 60.60.

There are no primers with Tm difference of <5.

5. Which 3 primer pairs have less self-complementary value?

- Primer 1 has a difference of 0
- Primer 4 has a difference of 0
- Primer 10 has a difference of 0