User:Crandel5425/sandbox



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, or Alexa Fluor dyes , 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
What we currently found:


 * 5-(3-Aminoallyl)-2’-Uridine (AA-UTP)
 * 5-(3-Aminoallyl)-2’-Deoxyuridine (AA-dUTP)
 * 5-(3-Aminoallyl)-2’-Cytidine (AA-CTP)
 * 5-(3-Aminoallyl)-2’-Deoxycytidine (AA-dCTP)
 * We should add images from the referenced webpage if possible. Crandel5425 (talk) 03:22, 12 March 2014 (UTC)

Use

 * 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. Mishasubz (talk) 09:48, 20 March 2014 (UTC)

Found information to expand article
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 Mishasubz (talk) 21:40, 22 March 2014 (UTC)