DNA基础知识

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Desalting Oligonucleotides by Gel FiltrationGeneralDesalting or buffer exchange on a gel filtration column is dependent on volume not quantity of DNA in solution. Many sizes of gel filtration columns ...

Recommended Storage Conditions Probes should be stored frozen both when lyopholized or in solution. Aliquoting your probe into additional vials after resuspension can help to minimize potential breakd ...

General Design GuidelinesThe following guidelines should be taken into account when designing modified oligonucleotides. Sequence Length - SYNTHEGEN can synthesize oligonucleotides from 5 to 110 bases ...

ROX StandardThe final working concentration of ROX Standard in the PCR buffer is 0.5 µMolar (1X). SYNTHEGEN's ROX Standards are shipped in liquid form at a concentration of 100 µMolar (200X). ...

1. Prior to precipitation dilute DNA solution may be concentrated to allow precipitation in smaller volumes.2. Add an equal volume of 2-Butanol to DNA solution and mix by vortexing. 3. Centrifuge samp ...

1. If the trityl group is not removed from a newly-synthesized oligonucleotide it may interfere with polymerization reactions.2. Evaporate the oligonucleotide to dryness.3. Add 300 ml 80% Acetic acid. ...

1. Pour and polymerize a 20% polyacrylamide gel no Urea.2. Remove clamps. Rinse with water. Remove comb. Rinse top of gel well.3. Insert comb teeth down until teeth just touch the gel. Mount the plate ...

How to Calculate the Melting Temperature of Oligonucleotides The melting temperature Tm of an oligonucleotide is its most critically important value. The most reliable and accurate determination of me ...

UV Quantitation of DNA DNA absorbs ultraviolet light due to its highly conjugated nature. DNA may thus be easily quantitated in a UV spectrometer. Typically 1 OD260 (i.e. a solution having an absorban ...

Oligonucleotide VisualizationThis is a question asked quite a bit most recently (late September 1995) in the bionet.molbio.methds-reagnts newsgroup. Here are two methods and who submitted them. UV Sha ...

UV shadowing is a technique for visualizing nucleic acids separated on acrylamide/urea gels. The technique utilizes shortwave UV light (254 nm) and a fluor-coated TLC plate. We recommend UV shadowing ...

In situ immunodetection of 5-methylcytosine (5-mC) on squashed cells using anti-methylcytosine Cross-references are to Schwarzacher & Heslop-Harrison 2000. Practical in situ Hybridization. Bios Oxfor ...

Methylation analyis using restriction enzyme digestionContributed by Dr. A. GratchevThis is a classical method of methylation analysis based on the property of some restriction enzymes to be unable to ...

DNA-Methyltransferase AssayThis protocol was written by Jean-Pierre Issa based on Adams et al.* Kam-Wing Jair has made some useful shortcuts that work well if you are careful.Here is Kam-Wing Jair's ...

DNA methylation is an epigenetic event that affects cell function by altering gene expression and refers to the covalent addition of a methyl group catalyzed by DNA methyltransferase (DNMT) to the 5-c ...

Calcium Phosphate Transfection of Neurons in Primary CultureDeveloped by Hank Dudek and Zhengui Xia (11/96)Modified by Steve Finkbeiner (rev. 7/97)Departments of Neurology and Physiology UCSF/GIND415- ...

CaPO4 Transfection for Chromaffin Cells1. 2M CaCl2 - 11.1g CaCl2 (anhydrous) + 47.3 ml H2O (or 14.7g CaCl2-2H2O) Filter sterilize and store at 4'C 2. 150 mM Na2HPO4 -2.13g Na2HPO4 + 99.7 ml H2O (or 4. ...

Materials:LipofectamineIMDM containing 10% fetal bovine serum 1% glutamine 1% aaIMDM containing 1% glutamineIMDM containing 20% fetal bovine serum 1% glutamine 1% aaIn a six-well or 35 mm tissue cultu ...