Summary

Nucleosome yoğunluk analiz için kitaplıklarına sıralama yerli Kromatin Immunoprecipitation nesil

Published: December 12, 2017
doi:
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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Disclosures

The authors have nothing to disclose.

Acknowledgements

Materials

1M Tris-HCl –pH 7.5Thermo Scientific15567-027
1M Tris-HCl –pH 8Thermo Scientific15568-025
0.5M EDTAThermo ScientificAM9260G
5M NaClSigma1001385276
Triton X-100Sigma1001412354
Sodium-DeoxycholateSigma1001437582
SDSThermo Scientific15525-017
Sodium-BicarbonateFisher ScientificS233-500
100% EtOHNANA
V-bottom 96 well plate (AB 1400)Thermo ScientificAB-1400-L
Gilson P2 pipetmanMandelF144801
Gilson P10 pipetmanMandelF144802
Gilson P20 pipetmanMandelF123600
Gilson P100 pipetmanMandelF123615
Gilson P200 pipetmanMandelF123601
Gilson P1000 pipetmanMandelF123603
Micrococcal Nuclease NEBM0247S
Thermo Mixer C (Heating block mixer)Eppendorf5382000023
Multi 12-channel Pippet P20Rainin17013803
Multi 12-channel Pippet P200Rainin17013805
1.5 ml LoBind tubeEppendorf22431021
0.5 ml LoBind tubeEppendorf22431005
Plastic Plate CoverEdge Bio48461
PCR coverBio RadMSD-1001
Aluminum Plate CoverBio RadMSF-1001
Elution buffer (EB buffer)Qiagen19086
1M DTTSigma646563
Eppendorf 5810R centrifuge Eppendorf22625004
Ultrapure waterThermo Scientific10977-015
Protein G dynabeadsThermo Scientific10001D
Protein A dynabeadsThermo Scientific10001D
Protease inhibitor CocktailCalbiochem539134
Rotating platformMandel1b109-12052010
Plate magnetAlpaqua2523
Domed 12-cap stripBio RadTCS1201
Buffer G2Qiagen1014636
ProteaseQiagen19155
Tube Magnet (DynaMag-2)Thermo Scientific12321D
Tube Magnet (DynaMag-2)LabCore730-006
V shape Plastic reservoirMandelS0100A
VortexMandelC1801
Mini FugeMettler ToledoXS2035
Analytical scaleMandelLB109-12052010
Quick Ligation Reaction Buffer, 5XNEBB6058S
DNA Polymerase I, Large (Klenow) FragmentNEBM0210S
Klenow Fragment (3'-5' exo)NEBM0212S
T4 DNA PolymeraseNEBM0203S
T4 Polynucleotide KinaseNEBM0201S
Deoxynucleotide Solution mix, 10mMNEBN0447S
dATP Solution 10mMNEBN0440S
Quick T4 DNA LigaseNEBM0202T
Adenosine 5'-Triphosphate (ATP), 25mMNEBP0756S
DNA PolymeraseThermo ScientificF549L
NEBuffer 2NEBB7002S
Hank's buffered salt solutionThermo Scientific14025076
Fetal bovine serum Thermo ScientificA3160702
phosphate buffer saline Thermo Scientific10010023
H3K4me3Cell Signaling9751S
H3K4me1DiagenodeC15410037
H3K27me3DiagenodepAb-069-050
H3K36me3Abcamab9050
H3K9me3DiagenodeC15410056
SeraMag  super-paramagnetic beads

References

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Generation of Native Chromatin Immunoprecipitation Sequencing Libraries for Nucleosome Density Analysis

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Cite This Article
Lorzadeh, A., Lopez Gutierrez, R., Jackson, L., Moksa, M., Hirst, M. Generation of Native Chromatin Immunoprecipitation Sequencing Libraries for Nucleosome Density Analysis. J. Vis. Exp. (130), e56085, doi:10.3791/56085 (2017).

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