| Parameter | Value | Context |
|---|---|---|
| EcoRI recognition sequence | 5'-GAATTC-3' | Palindromic hexanucleotide |
| EcoRI sticky end overhang | 4 nucleotides (AATT) | 5' single-stranded overhang |
| EcoRI source | E. coli strain RY13 | "Eco" = E. coli, "R" = RY13 |
| Restriction enzymes known | >900 | From 230+ bacterial strains |
| Denaturation temperature (PCR) | 94–98 °C | Breaks H-bonds between base pairs |
| Annealing temperature (PCR) | 50–65 °C | Primers bind template |
| Extension temperature (PCR) | 72 °C | Optimal for Taq polymerase |
| PCR amplification | 2ⁿ copies | n = number of cycles |
| 30 cycles of PCR | ~10^{9} (1 billion) copies | Exponential amplification |
| Heat shock temperature | 42 °C | For bacterial transformation |
| Heat shock duration | 60–90 seconds | After competency treatment |
| pBR322 insert capacity | Up to ~10 kb | Plasmid vector limit |
| Lambda phage insert capacity | Up to ~23 kb | Larger than plasmid vectors |
| EcoRI cuts at | Between G and A | On both strands in staggered manner |
| Taq polymerase source | Thermus aquaticus | Thermophile from hot springs |
| Gene gun particles | Gold or tungsten | Coated with DNA, fired at high velocity |
| Ethidium bromide emission | Orange under UV | Intercalates between DNA base pairs |
Memory anchors:
- 94-55-72: PCR temperatures (approx. annealing often given as 55 °C in NEET problems)
- 2ⁿ: PCR copy formula
- 42 °C: Heat shock for bacterial transformation
- GAATTC: EcoRI palindrome (6 letters, 6 nucleotides)