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Aminoallyl nucleotides or (AA-NTPs) contain a primary amine group on a linker that reacts with the amino-reactive dye such as a cyanine,HiLyte Fluor[1] , or Alexa Fluor dyes[2] , which contain a reactive leaving group, such as a succinimidyl ester (NHS). (base-pairing amino groups are not affected). These nucleotides are used for labeling DNA for example, 5-(3-Aminoallyl)-Uridine(AA-UTP) is more effective for high density labeling of DNA than pre-labeling DNA. Aminoallyl NTPs can be used for indirect DNA labeling in PCR, nick translation, primer extensions and cDNA synthesis. These labeled NTPs are helpful due to their application in molecular biology labs where radioactive labeling is not allowed or is banned.
Aminoallyl Nucleotides
editWhat we currently found:
- 5-(3-Aminoallyl)-2’-Uridine (AA-UTP)[3]
- 5-(3-Aminoallyl)-2’-Deoxyuridine (AA-dUTP)[3]
- 5-(3-Aminoallyl)-2’-Cytidine (AA-CTP)[3]
- 5-(3-Aminoallyl)-2’-Deoxycytidine (AA-dCTP) [3]
- We should add images from the referenced webpage if possible. Crandel5425 (talk) 03:22, 12 March 2014 (UTC)
Use
edit- cDNA (describe each in detail)
- PCR nick-translation
- random-primed labeling
- primer extension.
- DNA microarray, which measures the level of gene expression
Post-synthesis Post-synthesis labeling avoids the problems found in direct enzymatic incorporation of Cy-labeled dNTPs by generating probes with equal labeling effectiveness. With indirect labeling, amine-modified NTPs are incorporated during reverse transcription or aRNA amplification. Amino allyl-NTPs are incorporated with similar efficiency as unmodified NTPs during enzymatic reactions such as reverse transcription. (how do i link the from the reference list? #2) Mishasubz (talk) 22:04, 23 March 2014 (UTC)
- Biotin-labeling:
Biotinylation, a process by which biotin covalently attaches to a protein, used as a form of labeling to detect protein or molecule of interest. For nonradioactive labeling such as random priming, PCR labeling or nice translation Biotin-dUTPs works really well. Generally used for the efficient generation of biotinylated targets which can be captured with streptavidin coated solid phases or detected by streptavidin conjugates.[5] Mishasubz (talk) 09:48, 20 March 2014 (UTC)
Found information to expand article
edit[6] [7] [8] [9] [10] [11] [12]
fluorescent Cy dyes greatly enhanced the labelling efficiency, as analysed by spectrophotometry and fluorescent hybridisation signals. Indirect labelling using aa-UTP resulted in 2- to 3-fold higher degrees of labelling and fluorescent signals than labelling by direct incorporation of Cy-UTP [13] Mishasubz (talk) 21:40, 22 March 2014 (UTC)
References
edit- ^ AnaSpec. "HiLyte Fluor™ Dyes". Retrieved 12 March 2014.
- ^ life technologies. "Aminoallyl dUTP". Retrieved 12 March 2014.
- ^ a b c d ChemCyte. "Base Modified Nucleoside Triphosphates". ChemCyte. Retrieved 12March 2014.
{{cite web}}: Check date values in:|accessdate=(help) - ^ Ott Scheler, scheler (15 May 2009). "Fluorescent labeling of NASBA amplified tmRNA molecules for microarray applications". BMC Biotechnology.
{{cite journal}}: Unknown parameter|coauthors=ignored (|author=suggested) (help) - ^ http://www.roche-applied-science.com/shop/custom-biotech/products/biotin-16-dutp-3613112-1#tab2.
{{cite web}}: Missing or empty|title=(help) - ^ Berti, L.; Medintz, I. L.; Alessandrini, A.; Facci, P. (2009 Jun 10). "A one-pot functionalization strategy for immobilizing proteins onto linear dsDNA scaffolds". Nanotechnology. 20 (23): 235101. doi:10.1088/0957-4484/20/23/235101. PMID 19448298.
{{cite journal}}: Check date values in:|date=(help) - ^ Schoetzau, T.; Langner, J.; Moyroud, E.; Roehl, I.; Vonhoff, S.; Klussmann, S. (2003 Sep-Oct). "Aminomodified nucleobases: functionalized nucleoside triphosphates applicable for SELEX". Bioconjugate Chemistry. 14 (5): 919–26. doi:10.1021/bc0256547. PMID 13129394.
{{cite journal}}: Check date values in:|date=(help) - ^ Xiang, C. C.; Kozhich, O. A.; Chen, M.; Inman, J. M.; Phan, Q. N.; Chen, Y.; Brownstein, M. J. (2002 Jul). "Amine-modified random primers to label probes for DNA microarrays". Nature Biotechnology. 20 (7): 738–42. doi:10.1038/nb0702-738. PMID 12089562.
{{cite journal}}: Check date values in:|date=(help) - ^ Thermo. "Aminoallyl-dUTP". Retrieved 26 February 2014.
- ^ Life Technologies. "Amino Allyl Labeling for Array Analysis". Retrieved 26 February 2014.
- ^ Jena Bioscience. "Dye-labeled Uridine Nucleotides - Aminoallyl-dUTP". Retrieved 26 February 2014.
- ^ Kaposi-Novak, P.; Lee, J. S.; Mikaelyan, A.; Patel, V.; Thorgeirsson, S. S. (Oct 2004). "Oligonucleotide microarray analysis of aminoallyl-labeled cDNA targets from linear RNA amplification". BioTechniques. 37 (4): 580, 582–6, 588. doi:10.2144/04374ST02. PMID 15517970.
- ^ Hoen, PA (March 2003). "Fluorescent labelling of cRNA for microarray applications". Nucleic Acids Research. 31 (5): 20e–20. doi:10.1093/nar/gng020. PMC 149842. PMID 12595569.
{{cite journal}}: Unknown parameter|coauthors=ignored (|author=suggested) (help)