Strategies for covalent attachment of DNA to beads.

نویسندگان

  • Gali Steinberg
  • Katie Stromsborg
  • Lynette Thomas
  • David Barker
  • Chanfeng Zhao
چکیده

Several covalent attachment chemistries were tested for the immobilization of DNA onto glass beads. The comparison was based on the ability of these chemistries to produce derivatized beads that give good hybridization signals. Cyanuric chloride, isothiocyanate, nitrophenyl chloroformate, and hydrazone chemistries gave us the best (yet comparable) hybridization signals. We further characterized the cyanuric chloride method for the number of attachment sites, number of hybridizable sites, hybridization kinetics, effect of linker length on hybridization intensity and stability of the derivatized beads.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

End-specific covalent photo-dependent immobilisation of synthetic DNA to paramagnetic beads.

A novel approach for light-dependent covalent immobilisation of synthetic DNA oligomers to amino-coated paramagnetic beads is described. A hetero-bifunctional photo-reactive cross-linking chemical, 4-nitrophenyl 3-diazopyruvate, is applied to attach 5' amino-modified DNA to both silica and polystyrene paramagnetic beads. The coupling yields are comparable with similar methods in which no photo-...

متن کامل

A versatile approach to high-throughput microarrays using thiol-ene chemistry.

Microarray technology has become extremely useful in expediting the investigation of large libraries of materials in a variety of biomedical applications, such as in DNA chips, protein and cellular microarrays. In the development of cellular microarrays, traditional high-throughput printing strategies on stiff, glass substrates and non-covalent attachment methods are limiting. We have developed...

متن کامل

Stabilization of a raw-starch-digesting amylase by multipoint covalent attachment on glutaraldehyde-activated amberlite beads.

Raw-starch-digesting enzyme (RSDA) was immobilized on Amberlite beads by conjugation of glutaraldehyde/ polyglutaraldehyde (PG)-activated beads or by crosslinking. The effect of immobilization on enzyme stability and catalytic efficiency was evaluated. Immobilization conditions greatly influenced the immobilization efficiency. Optimum pH values shifted from pH 5 to 6 for spontaneous crosslinkin...

متن کامل

Comparison between different strategies of covalent attachment of DNA to glass surfaces to build DNA microarrays.

DNA microarray is a powerful tool allowing simultaneous detection of many different target molecules present in a sample. The efficiency of the array depends mainly on the sequence of the capture probes and the way they are attached to the support. The coupling procedure must be quick, covalent, and reproducible in order to be compatible with automatic spotting devices dispensing tiny drops of ...

متن کامل

DNA gyrase subunit stoichiometry and the covalent attachment of subunit A to DNA during DNA cleavage.

Escherichia coli DNA gyrase contains a 1:1 ratio of protomers coded by the genes gyrA and gyrB. This along with previous results shows that the enzyme has two copies of each protomer and thus a molecular weight of 400,000. Abortion of the gyrase reaction results in double-strand breakage of the DNA and covalent attachment of both gyrA protomers to the 5'-cut ends. We conclude that the gyrA prot...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Biopolymers

دوره 73 5  شماره 

صفحات  -

تاریخ انتشار 2004