• DNA标记

• 点击：    作者：   来源： 日期：2006-11-09    本站论坛

DNA标记（主要内容如下）

•   DNA Labeling by Nick Translation
•   Random Primed Labeling
•   End-Labeling
•   Purification of Labeled DNA
•   Non-isotopic Labeling
•   Others

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DNA Labeling by Nick Translation

§         (Ambros Lab)

§         (Worthington)

§        

§         (Promega)

§         (Life Technologies)

Random Primed Labeling

·         Random Primer Labeling (Karl Clark)
Prepare radioactively labeled DNA strands for the detection of target DNA or RNA sequences in usr/localious applications including Southern and northern blot hybridization. Detailed protocol for labeling, purification and determination of incorporation

·         Random Primed Labelling with P32 (Molecular Genetics Network Lab)

·         (Worthington)

·         (Crawford Lab)

·         (Amersham Pharmacia)

·         (Life Technologies)

·         (Fermentas)
Labeling of DNA by random oligonucleotide-primed synthesis

·         Probes (PMCI Research)

End-Labeling

·         (Amersham Pharmacia)
Labeling of  oligonucleotides with a single fluorescein residue at the 5' end

·         5' end labeling of oligonucleotides with gamma-³²P-ATP (Petra Klaff)
Labeling and purification method in great detail

·         (Promega)

·         (Lazo Lab)

·         End-labeling of DNA fragments
Single end or double ends, can also be used for RNA

·         Labeling of Oligonucleotides (PMCI Research)
Label oligo with polynucleotide kinase. 

·         Alkaline phosphatase labeling of oligos (Promega)
Provides useful guide for preparing phosphatase conjugated oligos

·         (Roe, OU)

·         (Molecular Genetics Network Lab)

·         (Amersham Pharmacia)

·         Fill in Labeling of DNA (Mike A. Dyer)

·         (Fermentas)
T4 polynucleotide kinase  labeling (ds and ss DNA or RNA; protruding, recessed and blunt 5'-ends) for (1)end labeling of DNA or RNA probes in DNA hybridization and DNA footprinting experiments, etc; (2)obtaining labeled molecular weight markers;(3)obtaining labeled molecular weight markers; (4)labelling of primers for sequencing; (5)addition of 5'-phosphates to oligonucleotides to allow subsequent ligation.

·         Oligonucleotide labeling with ³²P (Hahn Lab)
T4 polynucleotide kinase labeling 

Purification of Labeled DNA

§         (Petra Klaff)
Labeling and purification method (ethanol and CTAB in great detail

§         (Botwell Lab)
Purification of labeled oligos

·         (gopher://iubio.bio.indiana.edu/)
For purifying labeled probe

·         (Schimenti Lab)
Used to removed unincorporated nucleotides from labeling reactions.

·         (Hahn Lab)

·         (Life Technologies)
Protocol for PAGE Purification of Oligonucleotides Protocol for PAGE Purification of Oligonucleotides Oligonucleotides contaminated by significant amounts of aborted synthesis products (e.g., n-1,...

Non-isotopic Labeling

·         PCR DNA Biotinylation (KPL)
A rapid method for biotinylating DNA probes through incorporation of biotin-N₄-dCTP via a thermostable DNA polymerase in the polymerase chain reaction

·         Determination of efficiency of digoxigenin labelling (Molecular Genetics Network Lab)

·         Non-isotopic Labeling (Gottschling Lab)
Protocols for preparing DNA, RNA probe and molecular marker

Others

·         (Amersham)
Guidelines for preparation of radioactively-labeled PCR products for use as probes