Table 1: Enzymatic incorporation of fluorescent Nucleotides.

PCR: Polymerase chain reaction with Taq Polymerase; MDA-based WGA: Multiple displacement amplification (MDA)-based whole genome amplification (WGA) with Phi29 DNA Polymerase; NT: Nick Translation with DNAse I / DNA Polymerase I;
Primer Extension: Primer Extension with Klenow 3′-5′exo^–; 3′-End Labeling: Incorporation with Terminal deoxynucleotidyl Transferase (TdT); RT: Reverse Transcription with Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV RT)

¹: in-house tested
²: external reference (see datasheet)
n/a: no data available
– : not applicable

Nick translation is the method of choice for the generation of labeled DNA probes (100- 500 bp) from large templates such as genomic DNA, cosmids, BAC clones. It is based on the concerted activities of DNAse I (introduction of random single-strand breaks (“nicks”) into double-stranded DNA) and Polymerase I that incorporates labeled dUTPs instead of their natural counterpart dTTP at the “nicked” sites thus creating ideal in situ hybridization probes. There is no net DNA synthesis in contrast to PCR-based labeling approaches.

Biotin & Digoxigenin Nick Translation Labeling Kits are available as well.

Table 1: Enzymatic incorporation of Biotinylated Nucleotides.

PCR: Polymerase chain reaction with Taq Polymerase; NT: Nick Translation with DNAse I / DNA Polymerase I;
Primer Extension: Primer Extension with Klenow 3′-5′exo^–; 3′-End Labeling: Incorporation with Terminal deoxynucleotidyl Transferase (TdT); RT:: Reverse Transcription with Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV RT)

¹: in-house tested
²: external reference (see datasheet)
n/a: no data available
– : not applicable

Table 1: Enzymatic incorporation of Digoxigenin (DIG)-/(DNP)-modified nucleotides.

PCR: Polymerase chain reaction with Taq Polymerase; NT: Nick Translation with DNAse I / DNA Polymerase I;
Primer Extension: Primer Extension with Klenow 3′-5′exo^–; 3′-End Labeling: Incorporation with Terminal deoxynucleotidyl Transferase (TdT); RT:: Reverse Transcription with Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV RT)

Nick translation is the method of choice for the generation of labeled DNA probes (100- 500 bp) from large templates such as genomic DNA, cosmids, BAC clones. It is based on the concerted activities of DNAse I (introduction of random single-strand breaks (“nicks”) into double-stranded DNA) and Polymerase I that incorporates labeled dUTPs instead of their natural counterpart dTTP at the “nicked” sites thus creating ideal in situ hybridization probes. There is no net DNA synthesis in contrast to PCR-based labeling approaches.

The combination of (poly)-HRP-conjugated Biotin/Digoxigenin detection reagents with AF 488 (Alexa Fluor® 488)-, AF546 (Alexa Fluor® 546)- or AF594 (Alexa Fluor® 594)-labeled tyramide further increases detection sensitivity of up to 100-fold.

Fluorescent Nick Translation Labeling Kits are available as well.

CLICK-functionalized Nucleotides for DNA Labeling

¹: in-house tested
²: external reference (see datasheet)
n/a: no data available
– : not applicable

Amine-modified Nucleotides for DNA Labeling

¹: in-house tested
²: external reference (see datasheet)
n/a: no data available
– : not applicable

3′-End Oligonucleotide Labeling Reagents