1. Practical Contents DNA Extraction Gel Electrophoresis
Sequence Retrieval
Primer Designing
PCR
Real Time PCR or Q-PCR
Reverse Transcriptase PCR
Restriction Enzymes
Restriction Mapping via NEB Cutter
Gene Cloning
Southern Blotting
Northern Blotting
Western Blotting
CRISPR-Cas9 system

2. Explore NCBI for sequence retrieval
Record video for better understanding

3. Why Are Primers Important?
Primers are what gives PCR its SPECIFICITY!!!
Good primer design: PCR works great.
Bad primer design: PCR works terrible.

4. What is a primer?
A primer is a short strand of RNA or DNA (generally about bases) that serves as a starting point for DNA synthesis. It is required for DNA replication because the enzymes that catalyze this process, DNA polymerases, can only add new nucleotides to an existing strand of DNA.

5. PCR primers are not designed to:
Repeat regions
Regions that can form primer- dimers
Regions with low G/C content

6. General Primer Design Criteria
Target: Sequence of interest
Length: Usually bp
G/C content: 40-60%
End with 1-2 GC pairs, if possible
Avoid mismatches at terminal ends
Check with databases for specificity………..

7. Web Sites of Some Programs That Perform PCR Primer Design
Site Address
Primer3
AutoPrime
RTPrimerDB
PrimerBank
QPPD

8. BLAST In-Silico PCR BLAT
Primer Specificity…?
BLAST In-Silico PCR BLAT