2-Aminoethoxypropargyl ddGTP

Sanger method is one of the most reliable and earliest DNA sequencing methods. DNA is synthesized from four deoxynucleotide triphosphates (dNTPs). Each new nucleotide is added to the 3′ -OH group of the last dNTP added. Dideoxythymidine triphosphates (ddTTPs) can be added to the growing DNA strand but when it is, chain elongation stops because there is no 3′ -OH for the next nucleotide to be attached to. The DNA to be sequenced is prepared as a single strand. This template DNA is supplied with a mixture of dATP, dGTP, dCTP and dTTP in ample quantities. A mixture of all four dideoxynucleotides (ddATP, ddGTP, ddCTP and ddTTP), each present in limiting quantities and each labeled with a "tag" that fluoresces a different color, are added. Because all four normal nucleotides are present, chain elongation proceeds normally until, by chance, DNA polymerase inserts a ddNTP (instead of the normal dNTPs). If the ratio of normal nucleotide to the dideoxy versions is high enough, some DNA strands will succeed in adding several hundred nucleotides before insertion of the ddNTPs halts the process. At the end of the incubation period, the fragments are separated by length from longest to shortest. The resolution is so good that a difference of one nucleotide is enough to separate that strand from the next shorter and next longer strand. Each of the four ddNTPs fluoresces a different color when illuminated by a laser beam and an automatic scanner provides a printout of the sequence. These ddATP, ddGTP, ddCTP and ddTTP amine derivatives are the essential building blocks for developing Sanger sequencing reagents.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
17086	1 umoles	Price

Citations

View all 27 citations: Citation Explorer

Polyadenylated sequencing primers enable complete readability of PCR amplicons analyzed by dideoxynucleotide sequencing

Authors:

Beranek, M., Drastikova, M., Petera, J.

Journal:

Acta Medica (Hradec Kralove) (2012): 160-4

UV-induced bond modifications in thymine and thymine dideoxynucleotide: structural elucidation of isomers by differential mobility mass spectrometry

Authors:

St-Jacques, A., Anichina, J., Schneider, B. B., Covey, T. R., Bohme, D. K.

Journal:

Anal Chem (2010): 6163-7

Analysis of processivity of mungbean dideoxynucleotide-sensitive DNA polymerase and detection of the activity and expression of the enzyme in the meristematic and meiotic tissues and following DNA damaging agent

Authors:

Roy, S., Choudhury, S. R., Sengupta, D. N.

Journal:

Arch Biochem Biophys (2008): 55-65

A dideoxynucleotide-sensitive DNA polymerase activity characterized from endoreduplicating cells of mungbean (Vigna radiata L.) during ontogeny of cotyledons

Authors:

Roy, S., Sarkar, S. N., Singh, S. K., Sengupta, D. N.

Journal:

FEBS J (2007): 2005-23

Mechanism-based suppression of dideoxynucleotide resistance by K65R human immunodeficiency virus reverse transcriptase using an alpha-boranophosphate nucleoside analogue

Authors:

Selmi, B., Boretto, J., Sarfati, S. R., Guerreiro, C., Canard, B.

Journal:

J Biol Chem (2001): 48466-72

Physical properties

Molecular weight	587.31
Solvent	Water

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure

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