A laboratory guide for in vivo studies of DNA methylation by H. P. Saluz, J. P. Jost (auth.)

By H. P. Saluz, J. P. Jost (auth.)

Show description

Read Online or Download A laboratory guide for in vivo studies of DNA methylation and protein/DNA interactions PDF

Best cell biology books

Golgi Apparatus: Structure, Functions and Mechanisms (Cell Biology Research Progress)

The Golgi gear is an organelle present in so much eukaryotic cells. the first functionality of the Golgi equipment is to technique and package deal macromolecules, corresponding to proteins and lipids, after their synthesis and prior to they make their option to their vacation spot. This ebook offers topical examine information within the research of Golgi equipment, together with Golgi corporation and tension sensing; signalling pathways controlling mitotic Golgi breakdown in mammalian cells; the function of Golgi gear within the organic mechanisms of hypericin-mediated photodynamic remedy; the function of the Trans-Golgi community (TGN) within the sorting of nonenzymic lysosomal proteins; and the mechanisms related to the position of Golgi equipment alteration in neurological issues prompted by way of manganese.

Astrocytes : Wiring the Brain

Astrocytes play varied roles in imperative apprehensive approach (CNS) functionality and disorder, and the connections that the astrocyte makes with different cells of the mind are crucial for various vital neural projects. Bringing jointly contributions from foreign specialists on the best in their box, Astrocytes: Wiring the mind emphasizes mobile connections and surveys the most up-tp-date findings on astrocyte task.

Stem Cells: From Basic Research to Therapy, Volume 2: Tissue Homeostasis and Regeneration during Adulthood, Applications, Legislation and Ethics

The second one quantity within the Stem Cells sequence concentrates at the mechanisms of stem mobilephone regeneration within the grownup organism with a view in the direction of knowing how misplaced tissue should be changed in the course of maturity and getting older. the second one concentration of this quantity is on stem phone id and manipulation, together with purposes in uncomplicated examine, drugs, and undefined.

Advances in Protein Chemistry, 3

ADVANCES IN PROTEIN CHEMISTRY VOL three.

Additional info for A laboratory guide for in vivo studies of DNA methylation and protein/DNA interactions

Example text

Centrifuge as above. 5 rnl of cold nucIei buffer. 5 ml of cold nucIei buffer containing 1 % NP-40 and vortex weIl. > Keep suspension for 5 min on ice. > Centrifuge crude nuclei for 5 min at 4000 rpm (HB-4 Sorvall rotor or equivalent) at 4°C. 5 rnl of nuclei buffer and transfer the suspension into a 15-ml sterile Falcon conical tube. > Add an equal volume of 2 x proteinase K buffer containing 600 ~g of proteinase K/rnl. > Seal the Ud ofthe tube with paral"ilm amI incubate 111 Genomic Footprinting the tubes in a horizontal position under water with reciprocal shaking at 37°C overnight.

Put the suspension inlo an Erlenmeyer flask and incubate at 3TC for 5-10 min in a waterbath shaker. > Filter the digest with a nylon grid (1-mm-wide mesh) assembled into a funnel so that the filtration can be done into a glass beaker. > Rinse the filter with cold llank's solution. > Centrifuge cells at O'C at 300 x g (1500 rpm using a Sorvall HB-4 rotor or equivalenl) for 3 min. > Wash cells 3 times with ice cold Hank's solution as above. > If the pellet contains too many red blood cells, resuspend it in 10 ml ofred celllysis buffer (room temperature).

5 ml) add 30 fll of a 10x restrietion buffer (use the same buffer recommended by the manufacturer but without bovine serum albumin; the sterile filtered 10x buffer is stored at -20°C). Add the 15 flg of genomic DNA, mix gently by tapping the tube. Add sterile water up to a final volume of 300 fll per tube. Mix gently by tapping the tube ami add 45 units ofthe chosen restriction enzyme. Should restricLion enzymes with 4 bp recognition sequences be used, 150 units will be needed. Mix again as described above and give a shorl spin of a few seconds in a microfuge.

Download PDF sample

Rated 4.24 of 5 – based on 36 votes