DNA Probe Synthesis Services
The DNA probe is one of the most commonly used nucleic acid probes, It consists of single-stranded or double-stranded DNA with a length of tens to hundreds or even thousands of base pairs. Under proper pH, temperature and ionic strength, the DNA probe will hybridize with complementary unlabeled single-stranded DNA or RNA in the sample through molecular denaturation, renaturation and high-precision base pairing to form a double-stranded complex.
As an important nucleic acid probe, DNA probes are widely used in biomedical fields such as genetic engineering, molecular biology research, medical testing and biochemical analysis.
The types of DNA probes:
- Genomic probe (genomic probe): derived from the gene itself in the genome, it can be the entire sequence of the gene, or a segment of the gene.
- cDNA probe: A probe obtained by transcribing mRNA from the corresponding gene and then by reverse transcription.
- Oligonucleotide probe: synthetic in vitro DNA fragment with 20-50 bases complementary to gene sequence.
Creative Biolabs is a professional biotechnology service platform that provides outsourced oligonucleotide synthesis services. We have professional researchers and strict quality control, and can provide you with the above types of DNA probe synthesis.
Synthesis method of DNA probe:
- Nick translation:
The gap translation method is the most commonly used probe labeling method. The main components of the reaction system are DNase (DNase I), E. coli DNA polymerase 1 (DNA polymerase |), three deoxyribonucleotides triphosphates, and one isotope. Labeled nucleotides (such as dATP, dTTP, dCTP, P-dGTP).
Notched translation flow chart
- Random primer:
Random primer synthesis of double-stranded probe is to combine oligonucleotide primer with DNA template, under the action of Klenow enzyme, to synthesize DNA probe. The size, yield, and specific activity of the synthesized product depend on the amount of template, primer, dNTP and enzyme in the reaction. Usually the average length of the product is 400-600 nucleotides.
Random primer flow chart
- End-labelling:
The end labeling method does not label the DNA full-length, but only introduces a label at its 5 end or 3'end for partial labeling. This labeling method can obtain full-length DNA probes.
Advantages of DNA probe method:
- The DNA probe is polycloned in a plasmid vector, the preparation method is simple.
- Compared with RNA probes, DNA probes are not easily degraded, and generally can effectively inhibit DNase activity.
- The labeling methods of DNA probes are relatively mature and can be labeled with isotopes or non-isotopes. There are many methods to choose from.
References
- Kuchlyan Jagannath, Mori Mattia, et al. What makes thienoguanosine an outstanding fluorescent DNA probe? Journal of the American Chemical Society. 2020.
- Nguyen MinhVu H, Wiederhold Nathan P, et al. Spiromastigoides asexualis: a Phylogenetic Analysis and Evaluation as a Cause of False-Positive Blastomyces DNA Probe Testing. Journal of Clinical Microbiology. 2020.
*For Research Use Only. Not for use in diagnostic procedures.