What is the Difference Between Nick Translation and End Filling?

Nick translation and end filling are two techniques used in molecular biology, but they serve different purposes:

Nick Translation:

• Incorporates radiolabeled nucleotides into DNA.
• Used for labeling probes for hybridization to detect specific DNA sequences.
• Requires the use of 5' to 3' exonuclease activity.
• Synthesizes labeled probes based on the activities of DNase 1 and E. coli DNA polymerase 1 enzymes.

End Filling:

• Produces blunt fragments when the fragments have sticky ends (single-stranded overhangs).
• Used to create blunt fragments for ligation into vectors.
• Does not require the use of 5' to 3' exonuclease activity.

The key difference between nick translation and end filling is their purpose: nick translation is a process that creates labeled probes for hybridization, while end filling is a technique that creates blunt fragments for ligation into vectors. Both techniques are routinely performed in molecular research labs and have great importance in the field.

Comparative Table: Nick Translation vs End Filling

Here is a table comparing the differences between Nick Translation and End Filling:

Nick translation is used for labeling probes for hybridization in order to detect specific genes or sequences, while end filling is used to make blunted fragments from sticky end fragments for ligation into vectors.