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Phenol-chloroform DNA extraction from sand flies1. Crash Individual sand flies in 0.5 ml lysis buffer (50 mM NaCl 10 mM EDTA 50 mM Tris-HCl pH 8). 2. Incubate The sand fly homogenates with 100 ng/ml ...

1. Transfer cells to a 15 ml tube.2. Add 5 ml TBS- to flasks shake & pour into tubes.3. Spin down the cells @ 1K RPM for 5'.4. Resuspend in 1 ml TBS-.5. Transfer to 1.5 ml tubes.6. Spin down @ 14K RPM ...

Materials:phenol:chloroform (1:1) chloroformAdd an equal volume of buffer-saturated phenol:chloroform (1:1) to the DNA solution. Mix well. Most DNA solutions can be vortexed for 10 sec except for high ...

SolutionsGel StocksDiluent 5X Buffer 25% Acrylamide209 g Urea 209 g Urea 209 g Ureaup to 500 ml Q 250 ml 10X TBE 120.8 g Acrylamideup to 500 ml Q 4.1 g BISup to 500 ml Q2.5 M NH4 OAc19.2 g NH4 OAcup t ...

Materials:• 0.8 % agarose gel in 1x TAE• Digested DNA• Glass Milk• NaI solution• New WashProcedure:1) Run digested DNA out on agarose gel slowly (70 V on BioRad gel)2) Use lon ...

This is a modification of a salting out procedure as described by Miller et al. (1988) evaluated at the DNA Laboratory Medical School Malta. When analysed by spectrophotometry 95% of extracted genomic ...

Materials: RPMI 1640 medium fetal calf serum (FCS) 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents Best.-Nr. 295892) cell cuture flask Phythemaglutinin PHA-L (Seromed M 5030) CO2 cell cul ...

1. Pick single colony and inoculate 250 ml of LB broth containing 100 m g/l ampicillin or appropriate antibiotic. Shake at 250 RPM overnight.2. Centrifuge cells in a Sovall GSA (250 ml)or SLA-3000 (50 ...

Glass wool method: Run TAE agarose gel and cut the appropriate band out with a clean razor blade. Poke a small hole with hot needle on the bottom of an eppendorf tube and jam the hole with a little of ...

TE solution 10 mM Tris (pH to 7.5) 1 mM EDTA (pH to 8.0 to dissolve)2.Frozen agarose gel piece containing the desired DNA fragmentSupplies: 1.Micropipetter and tips 2.Microcentrifuge and tubes 3.Spatu ...

Sephadex G-50 spun column purification Spin column purification can be used to change buffers without a concomitant change in solution volume to remove protein contaminants or to purify plasmid DNA su ...

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macerate tissue in Eppendorf tube without butter at RT add 400 m l extraction buffer vortex for 4 sec leave sample at RT until other samples are ready ( 1 h) spin in microfuge for 1 min transfer 300 m ...

PEG Preparation of Plasmid Plasmid isolated by this procedure can be used routinely for electrophoretic analysis restriction endonuclease digestion and transformation of E. Coli. sequencing PCR and mo ...

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 absorbance of one unit at 260 nm ...

ECKDescriptionThis method was suggested in a BRL "Focus" article several years ago (Zeugin & Hartley 1985). It is a useful way to avoid coprecipitation of proteins and accumulation of salt in the fina ...

The principle and procedures of RLGS method was first described by Hatada et al. (1991) and its improvement was described by Asakawa (1996). Basically based on their procedures we have successfully ap ...

Salmon Sperm DNA1.Dissolve 1 g salmon sperm DNA in 100 ml H2 O. 2.Autoclave (20 minutes) and aliquot in 1 ml/tube 3.Store at -20℃. (common freezer for stock solutions) ...

'Phenol' as used is Analar grade. Phenol should be melted at 65℃8-hydroxyquinoline added to a final concentration of 0.1% and equilibrated three times with an equal volume of 1M Tris.HCl pH 7.0. The f ...

Sample DNAs should not be highly degraded nor contain PCR inhibitors such as high concentrations of heme or chelating agents. For each individual assayed 250 ng of genomic DNA are digested separately ...