Index: Volume 4 Number 1
Next Article: Improved mRNA Isolation

• efficient 1 hour procedure

• > 90% of DNA is biotinylated

• end label is 100% biotin leading to more efficient and sensitive applications

• non-isotopic assay to monitor probe labeling efficiency included

• labeled probes suitable for non-isotopic assays or for immobilization on streptavidin solid supports

The LABEL-IT™ 3' Biotin End Labeling Kit provides an effective and easy to use system for the rapid labeling of any oligonucleotide (including PCR and sequencing primers), or other DNA molecules at the 3' end with biotin-14-dCTP. The labeling reactions are carried out following a simple 60 minute protocol using terminal deoxynucleotidyl transferase. One to four biotinylated nucleotides are added, with the majority of oligo being converted to a labeled nucleic acid with mostly two and three additions. Greater than 90% of DNA is converted to biotinylated product. The biotin end label is free of unlabeled nucleotides, allowing for highly efficient and sensitive assays.

The labeled probes are suitable for use in non-isotopic Northern blots, Southern blots, dot blots, in situ hybridizations, colony hybridizations, and other hybridization-based assays. The 3' biotin end labeled DNA can also be used in solid phase-based nucleic acid applications, using the biotin as an anchor for binding to streptavidin solid supports. The LABEL-IT™ 3' Biotin End Labeling Kit includes enough reagents to perform 25 end labeling reactions. In addition to the reagents needed to perform the labeling reactions, the kit contains a non-isotopic assay for assessing the efficiency of the biotin incorporation into the DNA. This kit eliminates the need for purchasing costly biotin-modified oligonucleotides.

	Figure: Lane 1, 5'-32P-labeled M13 oligodeoxynucleotide (24'mer); Lane 2, the 5'-32P-labeled M13 oligo (40 pmol) was labeled with biotin-14-dCTP (0.25 nmol) following the LABEL-IT(tm) 3' Biotin End Labeling Kit protocol.

Next Article: Improved mRNA Isolation