What is the Difference Between Nick Translation and Primer Extension?

Nick translation and primer extension are two important techniques used in molecular biology, but they serve different purposes and involve different processes:

Nick Translation:

• This process creates labeled DNA probes for various hybridization reactions.
• It involves the synchronized activity of DNA polymerase I in removing the RNA primer and adding deoxyribonucleotides.
• A nick (gap) is formed between the DNA strands, which is sealed using a DNA ligase.
• Enzymes used: DNase I and DNA polymerase.
• Importance: Nick translation facilitates the marking of specific DNA sequences.

Primer Extension:

• This technique is used to detect a specific RNA transcript from a mixture and quantify its size and amount.
• It involves the use of reverse transcriptase to synthesize complementary DNA (cDNA) of the RNA sequence.
• Primer annealing and reverse transcription occur only when the specific RNA sequence is present in the sample.
• Enzyme used: Reverse transcriptase.
• Importance: Primer extension is useful for studying gene expression and quantifying the amount of mRNA present in a sample.

In summary, nick translation is a process that creates labeled DNA probes using DNA polymerase I, while primer extension is a technique for detecting and quantifying specific RNA transcripts using reverse transcriptase.

Comparative Table: Nick Translation vs Primer Extension

Here is a table comparing the differences between Nick Translation and Primer Extension:

Nick Translation is a process that generates labeled DNA probes for various hybridization reactions, using DNase 1 and DNA polymerase enzymes. On the other hand, Primer Extension is a technique used to find specific RNA or study gene expression, utilizing the reverse transcriptase enzyme.