DNA/cDNA Labeling

Fluorophores and haptens, are the most commonly used labels for the generation of non-radioactive DNA/cDNA probes. They are enzymatically introduced into DNA/cDNA via modified nucleotides that are incorporated as substitutes for their natural counterparts in either a one-step procedure (introduction of a fluorescent or hapten-modified nucleotide) or a two-step procedure (introduction of a reactive group carrying nucleotide and subsequent fluorescent or hapten labeling).

 

Label Labeling Kits Single Nucleotides
Fluorophore Fluorescent PCR Labeling Kits
Fluorescent Nick Translation Labeling Kits		 Fluorescent Nucleotides
Hapten Biotin & Digoxigenin PCR Labeling Kits
Biotin & Digoxigenin Nick Translation Labeling Kits
Biotin & Digoxigenin 3′-End Oligonucleotide Labeling Kits		 Biotinylated Nucleotides Digoxigenin-/DNP-modified Nucleotides
Amine Amine-modified Nucleotides
CLICK CLICK-functionalized Nucleotides
LNA LNA-functionalized Nucleotides → Nucleotides & Nucleosides
Nucleotide dependent
 Enzyme Labeling Reagent Kits (w/o labeled nucleotide)  
Non-radioactive Labeling of DNA and RNA
Labeling of in situ hybridization DNA probes
Hapten-modified Nucleotides for DNA/RNA Labelling

Table 1: Standard enzymatic procedures for the preparation of non-radioactive DNA/cDNA probes.

Template Method Labeled Probe Labeling site Enzyme
DNA PCR DNA random Thermophilic polymerase e.g. Taq Polymerase
MDA-based WGA Phi29 DNA Polymerase
Nick Translation DNAse I / DNA Polymerase I
Primer Extension Klenow Fragment 3’-> 5’ exo-, Taq Polymerase
Primer Extension 3’ OH Terminal Deoxynucleotidyl Transferase (TdT)
RNA Reverse Transcription cDNA random Reverse Transcriptase e.g. MMLV

Fluorescent DNA/cDNA Labeling

Fluorescent Nucleotides

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)

DNA Labeling cDNA Labeling
Emission color Nucleotide Cat. No. PCR NT Primer Extension 3’ End Labeling RT
dUTP
Blue AMCA-6-dUTP NU-803-AMCA ✔2 ✔2 n/a n/a n/a
DEAC-dUTP NU-803-DEAC – ✔1 n/a n/a n/a
dUTP-ATTO-425 NU-803-425 ✔1 ✔1,2 ✔1 ✔1 n/a
dUTP-XX-ATTO-425 NU-803-XX-425 ✔1 ✔1 n/a n/a n/a
Green dUTP-XX-AF488 NU-803-XX-AF488 ✔1 ✔1 n/a n/a n/a
dUTP-XX-ATTO-488 NU-803-XX-488 ✔1,2 ✔1,2 ✔1,2 ✔1,2 n/a
Fluorescein-12-dUTP NU-803-FAMX ✔1,2 ✔1,2 ✔1,2 ✔1,2 n/a
Rhodamine-12-dUTP NU-803-RHOX – ✔1,2 ✔1 ✔1,2 n/a
Yellow-Green dUTP-XX-ATTO-532 NU-803-532 ✔1 ✔1 ✔1 ✔1 n/a
Yellow dUTP-Cy3 NU-803-Cy3 ✔1,2 ✔1,2 ✔1 ✔1 ✔2
dUTP-XX-Cy3 NU-803-XX-CY3 ✔1 ✔1 ✔1 n/a ✔1
dUTP-XX-AF555 NU-803-XX-AF555 ✔1 ✔1 n/a n/a n/a
dUTP-ATTO-550 NU-803-550 – ✔1 ✔1 ✔1 n/a
Orange dUTP-Texas Red NU-803-TXR ✔1,2 ✔1,2 ✔2 n/a n/a
dUTP-XX-Texas Red NU-803-XX-TXR ✔1,2 ✔1,2 ✔2 n/a n/a
dUTP-XX-AF594 NU-803-XX-AF594 ✔1,2 ✔1,2 ✔1 n/a n/a
dUTP-XX-594 NU-803-XX-594 ✔1 ✔1 ✔1
Red dUTP-Cy5 NU-803-CY5 ✔1,2 ✔1,2 ✔1 n/a ✔2
dUTP-XX-Cy5 NU-803-XX-CY5 ✔1 ✔1 ✔1 n/a ✔1
dUTP-ATTO-647N NU-803-647N – ✔1,2 ✔1 ✔1 n/a
dUTP-ATTO-643 NU-803-643 ✔1 ✔1 ✔1
dUTP-XX-ATTO-643 NU-803-XX-643 ✔1 ✔1 ✔1
dUTP-XX-AF647 NU-803-XX-AF647 ✔1 ✔1 n/a n/a n/a
dUTP-ATTO-655 NU-803-655 – ✔1 ✔1 ✔1 n/a
dCTP
Green Fluorescein-12-dCTP NU-809-FAMX ✔1 ✔1 n/a n/a n/a
Rhodamine-12-dCTP NU-809-RHOX – ✔1 n/a n/a n/a
Yellow dCTP-Cy3 NU-809-Cy3 ✔1 ✔1 n/a n/a n/a
dCTP-ATTO-550 NU-809-550 – ✔1 n/a n/a n/a
Orange dCTP-Texas Red NU-809-TXR ✔1 ✔1 n/a n/a n/a
Red dCTP-Cy5 NU-809-CY5 ✔1 ✔1 n/a n/a n/a
dCTP-ATTO-647N NU-809-647N – ✔1 n/a n/a n/a

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

Non-radioactive Labeling of DNA and RNA
Labeling of in situ hybridization DNA probes
Products & Ordering
AMCA-6-dUTP NU-803-AMCA Aminoallyl-dUTP – AMCA DEAC-dUTP NU-803-DEAC Aminoallyl-dUTP – DEAC Aminoallyl-dUTP-XX-ATTO-488 NU-803-XX-488
Aminoallyl-dUTP-ATTO-425 NU-803-425 Aminoallyl-dUTP-XX-AF488 NU-803-XX-AF488 also known as Alexa Fluor 488®-dUTP Fluorescein-12-dUTP NU-803-FAMX Fluorescein-X-(5-aminoallyl)-dUTP
Aminoallyl-dUTP-XX-ATTO-425 NU-803-XX-425 Rhodamine-12-dUTP NU-803-RHOX Rhodamine-X-(5-aminoallyl)-dUTP Rho-green-X-dUTP, RhG-dUTP, R110-dUTP Aminoallyl-dUTP-XX-AF594 NU-803-XX-AF594 also known as Alexa Fluor 594®-dUTP
Aminoallyl-dUTP-XX-ATTO-532 NU-803-XX-532 Aminoallyl-dUTP-ATTO-550 NU-803-550 Aminoallyl-dUTP-XX-ATTO-594 NU-803-XX-594
Aminoallyl-dUTP-Cy3 NU-803-CY3 Aminoallyl-dUTP-Texas Red NU-803-TXR Aminoallyl-dUTP-Cy5 NU-803-CY5
Aminoallyl-dUTP-XX-Cy3 NU-803-XX-CY3 Aminoallyl-dUTP-XX-Texas Red NU-803-XX-TXR Aminoallyl-dUTP-XX-Cy5 NU-803-XX-CY5
Aminoallyl-dUTP-XX-AF555 NU-803-XX-AF555 Aminoallyl-dUTP-XX-ATTO-643 NU-803-XX-643 Aminoallyl-dUTP-ATTO-647N NU-803-647N
Fluorescein-12-dCTP NU-809-FAMX Fluorescein-X-5-Propargylamino-dCTP Aminoallyl-dUTP-XX-AF647 NU-803-XX-AF647 Aminoallyl-dUTP-ATTO-643 NU-803-643
Rhodamine-12-dCTP NU-809-RHOX Rhodamine-X-5-propargylamino-dCTP Rho-green-X-dCTP, RhG-dCTP, R110-dCTP Aminoallyl-dUTP-ATTO-655 NU-803-655 5-Propargylamino-dCTP-Cy3 NU-809-CY3
5-Propargylamino-dCTP-Texas Red NU-809-TXR 5-Propargylamino-dCTP-Cy5 NU-809-CY5 5-Propargylamino-dCTP-ATTO-550 NU-809-550
5-Propargylamino-dCTP-ATTO-647N NU-809-647N

Fluorescent PCR Labeling Kits

Labeled DNA probes e.g. for (Fluorescence) in situ hybridization((F)ISH are routinely prepared via PCR if template amounts are limited or amplification of specific DNA fragments is required.

We offer High Fidelity PCR Labeling Kits …

  • … for both fluorescent (e.g Cy®-, ATTOTM– or Alexa Fluor®-), Biotin and Digoxigenin labeling
  • … suitable for preparation of both short (>100 bp) and long DNA probes (up to 4 kbp tested) due to a High Fidelity Polymerase blend consisting of Taq polymerase and a proofreading enzyme.
  • … with convenient configurations for application-specific assay optimization such as single nucleotide format for optimization of labeled dUTP/dTTP ratio, positive control reaction (amplification of 500 bp fragment), testkits containing selected labeled dUTPs to find the optimal label
Fluorophore Selection Guide for PCR & Nick Translation Labeling Kits
Products & Ordering
HighFidelity Fluorescein PCR Labeling Kit APP-101-FAMX Preparation of Fluorescein-labeled DNA probes by PCR HighFidelity ATTO488 PCR Labeling Kit APP-101-488 Preparation of ATTO488-labeled DNA probes by PCR HighFidelity AF488 PCR Labeling Kit APP-101-AF488 Preparation of AF488-labeled DNA probes by PCR
HighFidelity GREEN PCR Labeling Testkit APP-101-GREEN Preparation of FAMX-, ATTO488- and AF488-labeled DNA probes by PCR HighFidelity Cy3 PCR Labeling Kit APP-101-CY3 Preparation of Cy3-labeled DNA probes by PCR HighFidelity AF555 PCR Labeling Kit APP-101-AF555 Preparation of AF555-labeled DNA probes by PCR
HighFidelity YELLOW PCR Labeling Testkit APP-101-YELLOW Preparation of Cy3- and AF555-labeled DNA probes by PCR HighFidelity AF594 PCR Labeling Kit APP-101-AF594 Preparation of AF594-labeled DNA probes by PCR HighFidelity ATTO594 PCR Labeling Kit APP-101-594 Preparation of ATTO594-labeled DNA probes by PCR
HighFidelity ORANGE PCR Labeling Testkit APP-101-ORANGE Preparation of AF594-, ATTO594- and TexasRed-labeled DNA probes by PCR HighFidelity Cy5 PCR Labeling Kit APP-101-CY5 Preparation of Cy5-labeled DNA probes by PCR HighFidelity AF647 PCR Labeling Kit APP-101-AF647 Preparation of AF647-labeled DNA probes by PCR
HighFidelity ATTO643 PCR Labeling Kit APP-101-643 Preparation of ATTO643-labeled DNA probes by PCR HighFidelity RED PCR Labeling Testkit APP-101-RED Preparation of Cy5-, AF647- and ATTO643-labeled DNA probes by PCR

Fluorescent Nick Translation Labeling Kits

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.

Fluorophore Selection Guide for DNA & RNA Labeling Kits
Products & Ordering
Atto425 NT Labeling Kit PP-305-425 Blue-green fluorescent DNA labeling by nick translation Fluorescein NT Labeling Kit PP-305-FAMX Green fluorescent DNA labeling by nick translation AF488 NT Labeling Kit PP-305-AF488 Green fluorescent DNA labeling by nick translation
Atto488 NT Labeling Kit PP-305-488 Green fluorescent DNA labeling by nick translation Cy3 NT Labeling Kit PP-305-CY3 Yellow fluorescent DNA labeling by nick translation Atto550 NT Labeling Kit PP-305-550 Yellow fluorescent DNA labeling by nick translation
AF555 NT Labeling Kit PP-305-AF555 Yellow fluorescent DNA labeling by nick translation TexasRed NT Labeling Kit PP-305-TXR orange DNA labeling by nick translation AF594 NT Labeling Kit PP-305-AF594 Orange fluorescent DNA labeling by nick translation
Atto647N NT Labeling Kit PP-305-647N Red fluorescent DNA labeling by nick translation AF647 NT Labeling Kit PP-305-AF647 Red fluorescent DNA labeling by nick translation Cy5 NT Labeling Kit PP-305-CY5 Red fluorescent DNA labeling by nick translation
Atto680 NT Labeling Kit PP-305-680 IR-red fluorescent DNA labeling by nick translation

Hapten Labeling of DNA/cDNA

Biotinylated Nucleotides

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)

DNA Labeling cDNA
Labeling
Nucleotide Cat. No. PCR NT Primer Extension 3’ End Labeling RT
dUTP
Biotin-11-dUTP C5 position NU-803-BIOX ✔1,2 ✔2 ✔2 ✔2 ✔2
Biotin-16-dUTP C5 position NU-803-BIO16 ✔1,2 ✔2 ✔2 ✔2 ✔2
dCTP
Biotin-11-dCTP C5 position NU-809-BIOX ✔1,2 ✔2 ✔2 ✔2 ✔2
Biotin-16-dCTP C5 position NU-809-BIO16 ✔1 ✔2 ✔2 ✔2 ✔2
Biotin-14-dCTP N4 position NU-956-BIO14 – ✔2 ✔2 n/a n/a
dATP
Biotin-7-dATP N6 position NU-835-BIO – ✔2 ✔2 ✔– n/a
Biotin-14-dATP N6 position NU-835-BIO14 – ✔2 ✔2 n/a n/a
Desthiobiotin-7-dATP N6 position NU-835-Desthiobio – n/a ✔2 n/a n/a
Biotin-11-dATP 7-Deaza position NU-1175-BIOX ✔1,2 ✔2 ✔2 n/a ✔2
dGTP
Biotin-11-dGTP 7-Deaza position NU-1615-BIOX n/a n/a n/a n/a n/a
ddUTP
Biotin-11-ddUTP C5 position NU-1619-BIOX – – – ✔2 –
ddATP
Biotin-11-ddATP 7-Deaza position NU-1612-BIOX – – – ✔2 –

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

Non-radioactive Labeling of DNA and RNA
Labeling of in situ hybridization DNA probes
Hapten-modified Nucleotides for DNA/RNA Labelling
Products & Ordering
Biotin-11-dUTP NU-803-BIOX Biotin-X-(5-aminoallyl)-dUTP Biotin-16-dUTP NU-803-BIO16 Biotin-16-(5-aminoallyl)-dUTP
Biotin-16-dCTP NU-809-BIO16 Biotin-16-Propargylamino-dCTP Biotin-7-dATP NU-835-BIO N6-(6-Aminohexyl)-dATP – Biotin
Biotin-11-dATP NU-1175-BIOX Biotin-11-ddUTP NU-1619-BIOX Biotin-X-5-Propargylamino-ddUTP
Biotin-14-dATP NU-835-BIO14 Biotin-11-ddATP NU-1612-BIOX
Biotin-14-dCTP NU-956-BIO14 Biotin-14-dCTP NU-956-BIO14
Biotin-11-dGTP NU-1615-BIOX Biotin-11-dGTP NU-1615-BIOX
D-Desthiobiotin-7-dATP NU-835-Desthiobio N6-(6-Aminohexyl)-dATP – D-Desthiobiotin Biotin-11-dCTP NU-809-BIOX Biotin-X-5-Propargylamino-dCTP

Digoxigenin-/DNP-modified Nucleotides

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)

DNA Labeling cDNA
Labeling
Nucleotide Cat. No. PCR NT Primer Extension 3’ End Labeling RT
dUTP
Digoxigenin-11-dUTP NU-803-DIGX ✔1,2 ✔1,2 ✔2 ✔2 ✔2
Aminoallyl-dUTP-DNP NU-803-DNP ✔1 n/a n/a n/a n/a
ddUTP
Digoxigenin-11-ddUTP NU-1619-DIGX – – – ✔2 –
Non-radioactive Labeling of DNA and RNA
Labeling of in situ hybridization DNA probes
Products & Ordering
DIG-11-dUTP NU-803-DIGX Aminoallyl-dUTP-DNP NU-803-DNP Aminoallyl-dUTP – Dinitrophenol, DNP-11-dUTP DIG-11-ddUTP NU-1619-DIGX

Biotin & Digoxigenin PCR Labeling Kits

Labeled DNA probes e.g. for in situ hybridization (ISH) are routinely prepared via PCR if template amounts are limited or amplification of specific DNA fragments is required.

We offer High Fidelity PCR Labeling Kits …

  • … for both Biotin & Digoxigenin and fluorescent labeling
  • … suitable for preparation of both short (>100 bp) and long DNA probes (up to 4 kbp tested) due to a High Fidelity Polymerase blend consisting of Taq polymerase and a proofreading enzyme.
  • … with convenient configurations for application-specific assay optimization such as single nucleotide format for optimization of labeled dUTP/dTTP ratio and a positive control reaction (amplification of 500 bp fragment).

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.

Products & Ordering
HighFidelity Biotin16 PCR Labeling Kit APP-101-BIO16 Preparation of Biotin16-labeled DNA probes by PCR HighFidelity Digoxigenin PCR Labeling Kit APP-101-DIGX Preparation of Digoxigenin-labeled DNA probes by PCR
AF488 tyramide reagent RNT-012 also known as Alexa Fluor® 488 tyramide AF546 tyramide reagent RNT-013 also known as Alexa Fluor® 546 tyramide
AF594 tyramide reagent RNT-014 also known as Alexa Fluor® 594 tyramide AF488 Streptavidin Conjugate RNT-015 Streptavidin, Alexa Fluor® 488 Conjugate
AF594 Streptavidin Conjugate RNT-016 Streptavidin, Alexa Fluor® 594 Conjugate

Biotin & Digoxigenin Nick Translation Labeling Kits

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.

Products & Ordering
Biotin16 NT Labeling Kit PP-310-BIO16 AF488 tyramide reagent RNT-012 also known as Alexa Fluor® 488 tyramide
Digoxigenin NT Labeling Kit PP-310-DIGX AF546 tyramide reagent RNT-013 also known as Alexa Fluor® 546 tyramide
AF594 tyramide reagent RNT-014 also known as Alexa Fluor® 594 tyramide AF488 Streptavidin Conjugate RNT-015 Streptavidin, Alexa Fluor® 488 Conjugate
AF594 Streptavidin Conjugate RNT-016 Streptavidin, Alexa Fluor® 594 Conjugate

Biotin & Digoxigenin 3' End Oligonucleotide Labeling Kits

3’-End labeled oligonucleotides can be used as sequence-specific protein binding or nucleic acid hybridization probes e.g. for EMSA, Northern or Southern blots. Compared to internal, random labeled probes, the label is located at the 3‘-End only and less likely interferes with probe binding.

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.

Figure 1 Terminal deoxynucleotidyl Transferase (TdT) template-independently transfers labeled nucleotides to the 3’-OH group of ssDNA/oligonucleotides. The number of nucleotide and thus label incorporation depends on the type of nucleotide ((dd)UTP) and type of label (n = 1 for labeled ddUTP, n = 1 – 3 (average) for labeled UTP, n > 1 for labeled dUTP (multiple label, tail length is highly nucleotide specific).

Labeling efficiency of BIO 3'-End Oligonucleotide Labeling Kit vs. competitor
KitLabel# of LabelLabeling principle similar to …
BIO 3‘-End Oligonucleotide Labeling Kit with Biotin-11-UTPBiotin1-3Biotin 3‘-End DNA Labeling Kit (Pierce/Thermo Scientific)
DIG 3‘-End Oligonucleotide Labeling Kit with Digoxigenin-11-ddUTPDigoxigenin1DIG Oligonucleotide 3‘ End Labeling Kit, 2nd generation (Roche)
3‘-End Oligonucleotide Labeling Reagent Kitnucleotide dependentnucleotide dependent
Products & Ordering
BIO 3′-End Oligonucleotide Labeling Kit with Biotin-11-UTP APP-001 AF546 tyramide reagent RNT-013 also known as Alexa Fluor® 546 tyramide
DIG 3′-End Oligonucleotide Labeling Kit with Digoxigenin-11-ddUTP APP-002 AF594 tyramide reagent RNT-014 also known as Alexa Fluor® 594 tyramide
3′-End Oligonucleotide Labeling Reagent Kit APP-003 AF488 Streptavidin Conjugate RNT-015 Streptavidin, Alexa Fluor® 488 Conjugate
AF488 tyramide reagent RNT-012 also known as Alexa Fluor® 488 tyramide AF594 Streptavidin Conjugate RNT-016 Streptavidin, Alexa Fluor® 594 Conjugate

Click Chemistry-based DNA/cDNA Labeling

CLICK-functionalized Nucleotides for DNA Labeling

Table 1: Enzymatic incorporation of CLICK-functionalized nucleotides.

PCR: Polymerase chain reaction with Taq Polymerase (CLK-T09, NU-1705) or family B polymerases Pwo, Deep Vent exo– and KOD XL (CLK-T05/-T06/-T07); NT: Nick Translation with DNAse I / DNA Polymerase I;
Primer Extension: Primer Extension with family B polymerases Pwo, Deep Vent exo– and KOD XL; 3′-End Labeling: Incorporation with Terminal deoxynucleotidyl Transferase (TdT); RT:: Reverse Transcription with Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV RT)

NucleotideCat. No.PCRNTPrimer Extension3’ End LabelingRT
dUTP
5-DBCO-PEG4-dUTPCLK-048✔1n/an/an/an/a
5-Azidomethyl-dUTPCLK-084n/an/an/an/an/a
Azide-PEG4-Aminoallyl-dUTPNU-1705✔1n/an/a✔2n/a
5-Ethynyl-dUTPCLK-T07✔1,2n/a✔2n/an/a
C8-Alkyne-dUTPCLK-T05✔1,2n/a✔2n/an/a
5-TCO-PEG4-dUTPCLK-035✔1n/an/an/an/a
5-Vinyl-dUTPCLK-068✔1n/an/an/an/a
dCTP
C8-Alkyne-dCTPCLK-T06✔1,2n/a✔2✔2n/a
5-DBCO-PEG4-dCTPCLK-060✔1n/an/an/an/a
5-Azido-PEG4-dCTPCLK-070✔1n/an/an/an/a
dATP
N6-Azidohexyl-dATPCLK-NU-002✔1n/an/an/an/a

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

Non-radioactive Labeling of DNA and RNA
Labeling of in situ hybridization DNA probes
Click Chemistry background information
Products & Ordering
5-DBCO-PEG4-dUTP CLK-048 5-Azidomethyl-dUTP CLK-084
Azide-PEG4-aminoallyl-dUTP NU-1705 5-Ethynyl-dUTP (5-EdUTP) CLK-T07
C8-Alkyne-dUTP CLK-T05 5-TCO-PEG4-dUTP CLK-035 5-trans-Cyclooctene-PEG4-dUTP
5-Vinyl-dUTP CLK-068 C8-Alkyne-dCTP CLK-T06
5-DBCO-PEG4-dCTP CLK-060 5-Azido-PEG4-dCTP CLK-070
N6-(6-Azido)hexyl-dATP CLK-NU-002

Amine Labeling of DNA/cDNA

Amine-modified Nucleotides for DNA Labeling

Table 1: Enzymatic incorporation of Amine-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)

NucleotideCat. No.PCRNTPrimer Extension3’ End LabelingRT
dUTP
5-Aminoallyl-dUTPNU-803✔1,2✔2✔2✔2✔2
dCTP
5-Propargylamino-dCTPNU-809✔1n/an/an/an/a
dATP
N6-6-Aminohexyl-dATP N6 positionNU-835-✔2n/an/an/a
7-Deaza-7-Propargylamino-dATP 7-Deaza positionNU-1611✔2n/an/an/an/a

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

Non-radioactive Labeling of DNA and RNA
Labeling of in situ hybridization DNA probes
Products & Ordering
Aminoallyl-dUTP – Solution NU-803 5-Propargylamino-dCTP – Solid NU-809-lyo
Aminoallyl-dUTP – Solid NU-803-LYO N6-(6-Aminohexyl)-dATP NU-835
5-Propargylamino-dUTP NU-998 7-Deaza-7-propargylamino-dATP NU-1611
5-Propargylamino-dCTP – Solution NU-809

LNA Labeling of DNA/cDNA

Products & Ordering
HighFidelity LNA PCR Labeling Kit APP-101-LNA Preparation of LNA-labeled DNA probes by PCR

Enzyme Labeling Reagent Kits for DNA/cDNA Labeling

3′-End Oligonucleotide Labeling Reagents

Products & Ordering
3′-End Oligonucleotide Labeling Reagent Kit APP-003

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