Mismatched Primer Extension Assays
نویسندگان
چکیده
منابع مشابه
Comparison of DNA polymerases for quantification of single nucleotide differences by primer extension assays.
en, depending on the rotisserie motor kit you choose. Widely available power bars can serve as an on/off switch for motors not equipped with their own. Membranes are inserted into the 50 mL tubes. The surface carrying the nucleic acids or proteins should face inwards. Membranes should be moist to prevent breakage and air bubble formation from surface tension. To start the operation, reagent sol...
متن کاملPredicting the success of primer extension genotyping assays using statistical modeling.
Using an empirical panel of more than 20 000 single base primer extension (SNP-IT) assays we have developed a set of statistical scores for evaluating and rank ordering various parameters of the SNP-IT reaction to facilitate high-throughput assay primer design with improved likelihood of success. Each score predicts either signal magnitude from primer extension or signal noise caused by misprim...
متن کاملAnalysis of RNA by primer extension.
1. Phosphorylate oligonucleotide probe: a. Oligonucleotide primer (5 – 7 pmoles or 60 ng) 1 μL b. dH2O 6.5 μL c. 10X kinase buffer 1.5 μL d. T4 PNK (~10 U) 1 μL e. [γ-P]ATP (7000 Ci/mmole) 2 μL 2. Stop the kinase reaction by adding 500 μL of TE (pH 7.6) 3. Add 25 μg of carrier RNA. 4. Phenol/Chloroform extract 2X 5. Ethanol precipitate with sodium acetate (pH 5.2) 6. Wash with 70% ethanol and r...
متن کاملHuman DNA polymerase kappa forms nonproductive complexes with matched primer termini but not with mismatched primer termini.
Human DNA polymerase kappa (pol kappa) is a member of the Y family of DNA polymerases that function in translesion synthesis. It synthesizes DNA with moderate fidelity and does not efficiently incorporate nucleotides opposite DNA lesions. Pol kappa has the unusual ability to efficiently extend from mismatched primer termini, and it extends readily from nucleotides inserted by other DNA polymera...
متن کاملEffect of terminal 3'-hydroxymethyl modification of an RNA primer on nonenzymatic primer extension.
The significance of the precise position of the hydroxyl at the 3'-end of an RNA primer for nonenzymatic template-directed primer extension is not well understood. We show that an RNA primer terminating in 3'-hydroxymethyl-2',3'-dideoxy-guanosine has greatly diminished activity, suggesting that the spatial preorganization of the terminal sugar contributes significantly to the efficiency of prim...
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ژورنال
عنوان ژورنال: BIO-PROTOCOL
سال: 2015
ISSN: 2331-8325
DOI: 10.21769/bioprotoc.1508