DNA基础知识

DESCRIPTIONLabeling of DNA by random oligonucleotide-primed synthesis is based on the investigations of A.Feinberg and B.Vogelstein 1 2 and is a good alternative to nick translation for producing uni ...

RADIOLABELING OF PROBES FOR ELECTROPHORETIC MOBILITY SHIFT ASSAYS ...

DNA probes are prepared using a modification of the method of Feinberg and Vogelstein (1983). You will need: Nuclease-free BSA-dCTP (Amersham or DuPont)Klenow DNA Polymerase (New England Biolabs or Ph ...

DNA probes are prepared using a modification of the method of Feinberg and Vogelstein (1983). You will need: Nuclease-free BSA-dCTP (Amersham or DuPont)Klenow DNA Polymerase (New England Biolabs or Ph ...

IntroductionThe efficiency of nucleotide incorporation in DNA/RNA polymerization reactions (e.g. transcription reverse transcription and DNA replication) can be determined by trichloroacetic acid (TCA ...

IntroductionThe efficiency of nucleotide incorporation in DNA/RNA polymerization reactions (e.g. transcription reverse transcription and DNA replication) can be determined by trichloroacetic acid (TCA ...

Prepare a ligation mix:Ligation Mix (2x)10x ligase buffer1.0 mldigested vector(0.1 mg/ml)1.0 mlH2O6.0 mltotal8.0 mldivide ligation mix into two Eppendorf tubesLigation Rxn InsertControlligation mix ...

Cohesive End Ligation1) The ligation mixture contains the following: vector DNA (~100 ng) insert DNA (equimolar or 2 or 3 X molar concentration of vector) water added to 18 µl 2) Heat the mixture at 4 ...

set up the following reaction:CIP Rxn H2O7.8 ml10x cip rxn buffer2.0 mlDNA(e.g; 3 kb vector; 0.2 mg/ml; 2 mg total)10.0 ml(1 u/ml) CIP0.2 mltotal20.0 mlincubate for 60 min at 37°C add 1.14 ml 0.2 M ...

DEPHOSPHORYLATION OF LINEARIZED DNAALKALINE PHOSPATASE(CIP) DIGESTION Digestion of protruding 5'-ends1. Precipitate digested DNA and resuspend in a 100ml of 1X CIP Buffer.2. Add 1U CIP for 100pmol 5'- ...

DEPHOSPHORYLATION OF LINEARIZED DNAALKALINE PHOSPATASE(CIP) DIGESTION Digestion of protruding 5'-ends1. Precipitate digested DNA and resuspend in a 100ml of 1X CIP Buffer.2. Add 1U CIP for 100pmol 5'- ...

Phosphatasing with Shrimp Alkaline Phosphatase (S.A.P.)Use to prevent linearized vector plasmid from recircularizing. The phosphatase from shrimp is easier to get rid of than is CIP because SAP is hea ...

Template PreparationThe quality of sequencing results is directlyrelated to the quality of the template. ABI recommends a minialkaline-lysis/PEG precipitation procedure (the Core can supplyinformation ...

Interpreting DNA Sequencing ResultsEvaluating ChromatogramsMany problems with sequencing results are not recognized by viewing the text file alone. Therefore the quality of your sequence should alway ...

The following dye-labeled terminator reaction chemistries have been designed to balance conservation of reagents with the resulting sequence product signal strength. When coupled with BACPAC Resource ...

Preparation of DNA Template For Direct Sequencing of Large Insert PAC and BACPlasmidundefined This protocol is designed to allow the use of 96-well trays (8x12 microtiter tray format) and multi-channel de ...

Preparation of DNA Template For Direct Sequencing of Large Insert PAC and BACPlasmidundefined This protocol is designed to allow the use of 96-well trays (8x12 microtiter tray format) and multi-channel de ...

MATERIALS: 2-glass plates 1 sharks -tooth comb and spacers Whatman 3 mm paper 30 or 40% acrylamide-bis (19:1) 10X TBE urea 10% ammonium persulfate TEMED 60 cc. syringe Rain-ex Preparation of glass pla ...

MATERIALS: 2-glass plates 1 sharks -tooth comb and spacers Whatman 3 mm paper 30 or 40% acrylamide-bis (19:1) 10X TBE urea 10% ammonium persulfate TEMED 60 cc. syringe Rain-ex Preparation of glass pla ...

DNA Sequencing GelsBuffers and gel solutionsLong Ranger: we started using this in early 1995. Great stuff; the best thing is that the gels are not sticky after drying even without removal of the urea. ...