Primer Design of LDHA (Lactate Dehydrogenase A)

Prepared by: Yasmine Osama Abdul hameed Supervised by: Dr. Heba Alwasify  The Steps: 1. I chose the nucleotide from the data base and then wrote the name of my gene in the search: Homo sapiens lactate dehydrogenase A (LDHA) [org]. Visual Learning Auditory learning 2. I chose Ho...

Prepared by: Yasmine Osama Abdul hameed Supervised by: Dr. Heba Alwasify  The Steps: 1. I chose the nucleotide from the data base and then wrote the name of my gene in the search: Homo sapiens lactate dehydrogenase A (LDHA) [org]. Visual Learning Auditory learning 2. I chose Homo sapiens lactate dehydrogenase A (LDHA), transcript variant 1, mRNA 3. I opened the (idtd) website and registered a new account on it.  4. Select Primer Quest Tool from Tools. 5. I clicked on the Download sequence link (s) using GenBank or Accession ID and then download the NCBI ID#..6I chose the appropriate design so that it is. Amplicon Length: 115, which is greater than 100. Then I clicked (View Assay Details) Steps to verify the correctness of the primer design: The first method: 7. I pressed the Hairpin button for both Forward and Reverse to make sure that the delta G is greater than -9. The second method: 8. Select Oligo Analyzer Tool from Tools. 9. I copied the Forward sequence and pasted it into Homo-Dimer Analysis to make sure that the delta G is greater than -9. Then I copied the Reverse sequence and pasted it into Hetero-Dimer Analysis for the same reason as well. Third method: 10. I went to the primer Blast website and copied both the Forward sequence and Reverse sequence into it and modified the Primer Pair Specificity Checking Parameters to ensure that there was no mismatching. Fourth method: 11. I went to the PCR Primer Stats website and copied both the Forward sequence and Reverse sequence and sent their data to make sure there was no warning or error.