Atjaunināt sīkdatņu piekrišanu

E-grāmata: Analysis of RNA-Protein Complexes in vitro

Edited by (Aarhus University, Aarhus, Denmark), Edited by (University of Copenhagen, Denmark), Edited by (Aarhus University, Aarhus, Denmark)
Citas grāmatas par šo tēmu:
  • Formāts - PDF+DRM
  • Cena: 202,25 €*
  • * ši ir gala cena, t.i., netiek piemērotas nekādas papildus atlaides
  • Ielikt grozā
  • Pievienot vēlmju sarakstam
  • Šī e-grāmata paredzēta tikai personīgai lietošanai. E-grāmatas nav iespējams atgriezt un nauda par iegādātajām e-grāmatām netiek atmaksāta.
Analysis of RNA-Protein Complexes in vitroPalielināt
Citas grāmatas par šo tēmu:

DRM restrictions

  • Kopēšana (kopēt/ievietot):

    nav atļauts

  • Drukāšana:

    nav atļauts

  • Lietošana:

    Digitālo tiesību pārvaldība (Digital Rights Management (DRM))
    Izdevējs ir piegādājis šo grāmatu šifrētā veidā, kas nozīmē, ka jums ir jāinstalē bezmaksas programmatūra, lai to atbloķētu un lasītu. Lai lasītu šo e-grāmatu, jums ir jāizveido Adobe ID. Vairāk informācijas šeit. E-grāmatu var lasīt un lejupielādēt līdz 6 ierīcēm (vienam lietotājam ar vienu un to pašu Adobe ID).

    Nepieciešamā programmatūra
    Lai lasītu šo e-grāmatu mobilajā ierīcē (tālrunī vai planšetdatorā), jums būs jāinstalē šī bezmaksas lietotne: PocketBook Reader (iOS / Android)

    Lai lejupielādētu un lasītu šo e-grāmatu datorā vai Mac datorā, jums ir nepieciešamid Adobe Digital Editions (šī ir bezmaksas lietotne, kas īpaši izstrādāta e-grāmatām. Tā nav tas pats, kas Adobe Reader, kas, iespējams, jau ir jūsu datorā.)

    Jūs nevarat lasīt šo e-grāmatu, izmantojot Amazon Kindle.

The central role of RNA in many cellular processes, in biotechnology, and as pharmaceutical agents, has created an interest in experimental methods applied to RNA molecules. This book provides scientists with a comprehensive collection of thoroughly tested up-to-date manuals for investigating RNA-protein complexes in vitro. The protocols can be performed by researchers trained in standard molecular biological techniques and require a minimum of specialized equipment. The procedures include recommendation of suppliers of reagents.
Preface v
Chapter
1. Introduction to RNA-protein interactions
1(12)
1.1. Introduction
1(1)
1.2. RNA secondary structures
1(4)
1.2.1. Helices
1(1)
1.2.2. Bulges and internal loops
2(2)
1.2.3. Hairpin loops
4(1)
1.2.4. Single-stranded regions
5(1)
1.3. RNA tertiary structures
5(1)
1.3.1. Triple-base pairs (single-strand/helix interactions)
5(1)
1.3.2. Pseudoknots (single-strand/single-strand interaction)
5(1)
1.3.3. Hairpin/internal loop interactions
6(1)
1.4. Protein motifs involved in RNA binding
6(5)
1.4.1. Ribonucleoprotein (RNP) motif
6(2)
1.4.2. The K homology domain (KH)
8(1)
1.4.3. The arginine-rich motif (ARM)
9(1)
1.4.4. The RGG box
9(1)
1.4.5. Zinc fingers
10(1)
1.4.6. Double-strand RNA binding domains (dsRBD)
10(1)
References
11(2)
Chapter
2. Preparation of RNA
13(44)
2.1. Working with RNA
13(6)
2.1.1. Special precautions when working with RNA
13(1)
2.1.2. Basic protocols for RNA work
14(1)
2.1.2.1. RNA quantification
14(1)
2.1.2.2. Gel purification
15(1)
2.1.2.3. Extraction with organic solvents
16(1)
2.1.2.4. Precipitation
16(1)
2.1.2.5. Storage of RNA
17(1)
2.1.2.6. RNA renaturation
17(1)
2.1.2.7. Desalting and removal of nucleotides
18(1)
2.1.2.8. Diethylpyrocarbonate
18(1)
2.1.2.9. Standard reagents
18(1)
2.2. Isolation of RNA from cells
19(13)
2.2.1. Guanidinium thiocyanate-CsCl method
20(3)
2.2.2. Single step guanidinium thiocyanate acid-phenol method
23(2)
2.2.3. Hot phenol SDS method
25(2)
2.2.4. Vanadyl ribonucleoside complex method for preparation of nuclear and cytoplasmic RNA
27(2)
2.2.5. Purification of poly-A^(+) RNA
29(3)
2.3. In vitro synthesis of RNA
32(21)
2.3.1. In vitro transcription of RNA
33(2)
2.3.1.1. Quantification of in vitro transcribed RNA (in the presence of trace amount of Alpha-[ (32)P] UTP)
35(1)
2.3.2. Co-transcriptional labelling or modification of RNA
35
2.3.2.1. Generation of specific terminus by RNase H
41
2.3.2.2. Cotranscriptional incorporation of nucleotides containing modified ribose moieties
40(3)
2.3.3. End-labelling of RNA
43(1)
2.3.3.1. Dephosphorylation of RNA
43(2)
2.3.3.2. 5'-end labeling of RNA
45(1)
2.3.3.3. 3'-end labeling of RNA
46(1)
2.3.3.3.1. RNA ligase method
46(1)
2.3.3.3.2. Poly (A) polymerase method
46(1)
2.3.3.4. Preparation of [ Alpha (32)P] p^Cp from Cp and [ Gamma-(32)P] ATP
47(1)
2.3.4. Specific modification of RNA at an internal site
48(5)
References
53(4)
Chapter
3. Preparation of protein
57(26)
3.1. Isolation of protein from cells
57(8)
3.1.1. Preparation of HeLa cell extract
57(5)
3.1.2. Purification of SR proteins from nuclear extract
62(3)
3.2. Expression and purification of recombinant proteins in E. coli
65(11)
3.2.1. Expression of recombinant proteins in E. coli
66(3)
3.2.2. Purification of GST-tagged proteins under native conditions
69(3)
3.2.3. Purification of His-tagged proteins under denaturing conditions
72(2)
3.2.4. Labeling of protein using heart muscle kinase
74(2)
3.3. In vitro translation
76(3)
References
79(4)
Chapter
4. Preparation and analysis of RNA-protein complexes in vitro
83(98)
4.1. Formation of RNA-protein complexes
83(1)
4.2. Analysis of RNA-protein complexes
84(12)
4.2.1. Filter binding assay
85(3)
4.2.2. Mobility shift analysis
88(4)
4.2.3. Sucrose gradient analysis
92(4)
4.3. Isolation and identification
96(14)
4.3.1. Affinity purification using tagged RNA or protein
96(3)
4.3.1.1. Binding of biotinylated protein or RNA to streptavidin beads
99(2)
4.3.1.2. Binding of protein to glutathione beads
101(1)
4.3.1.3. Affinity purification of RNA-protein complexes
102(3)
4.3.2. UV cross-linking
105(5)
4.4. RNA footprinting
110(37)
4.4.1. Enzymatic probing
114(3)
4.4.2. Chemical probing
117(2)
4.4.2.1. DMS (dimethylsulphate) modification for probing adenosines and cytidines
119(3)
4.4.2.2. DMS (dimethylsulphate) modification for probing guanosines
122(1)
4.4.2.3. DEP (diethylpyrocarbonate) modification for probing adenosines
123(2)
4.4.2.4. Kethoxal (3-ethoxy-2-oxo-butanal) modification for probing guanosines
125(1)
4.4.2.5. CMCT (1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide for probing uridines and guanosines
126(1)
4.4.2.6. Hydroxyl radical modification for probing ribose moieties
127(2)
4.4.2.7. Iodine scission of phosphorothioate containing RNA for probing phosphates
129(1)
4.4.3. Identification of modified residues
130(1)
4.4.3.1. Reverse transcriptase assay
130(3)
4.4.3.1.1. Extension from an end-labelled primer
133(1)
4.4.3.1.2. Extension in the presence of radioactive dATP
134(2)
4.4.3.1.3. Sequencing by the reverse transcriptase method
136(1)
4.4.3.2. End-labelling method
137(2)
4.4.3.2.1. Enzymatic sequencing of end-labelled RNA
139(3)
4.4.3.2.2. Chemical sequencing of end-labelled RNA
142(5)
4.5. Protein footprinting
147(14)
4.5.1. Protein footprinting using proteinases
150(7)
4.5.2. Protein footprinting using Fe^(2+)/H(2)O(2)
157(2)
4.5.3. Preparation of Tris/Tricine-SDS-polyacrylamide gels
159(2)
4.6. Modification interference
161(8)
4.6.1. Base modifications
163(1)
4.6.2. Phosphate modification by ethylnitrosourea (ENU)
164(4)
4.6.3. Analogue interference
168(1)
4.7. SELEX
169(7)
References
176(5)
Chapter
5. Functional analysis of RNA-protein complexes in vitro
181
5.1. Pre-mRNA splicing
181(23)
5.1.1. In vitro mRNA splicing
182(3)
5.1.2. Analysis of RNA splicing products by denaturing gel electrophoresis
185(4)
5.1.3. Analysis of RNA splicing complexes by native gel electrophoresis
189(2)
5.1.4. Debranching of RNA lariats
191(2)
5.1.5. Analysis of RNA splicing complexes by sucrose gradients
193(4)
5.1.6. Affinity purification and characterization of splicosome complexes
197(7)
5.2. Polyadenylation
204(6)
5.2.1. In vitro polyadenylation
204(3)
5.2.2. Analysis of mRNA polyadenylation states
207(3)
5.3. RNA modification
210(10)
5.3.1. Reverse transcriptase-polymerase chain reaction (RT-PCR)
213(5)
5.3.2. Primer extension by Klenow polymerase (after RT-PCR)
218(1)
5.3.3. Primer extension by reverse transcriptase
219(1)
5.4. Translation
220(11)
5.4.1. Preparation of ribosomes
220(1)
5.4.1.1. Bacterial ribosomes
221(2)
5.4.1.2. Mammalian ribosomes
223(2)
5.4.2. Preparation and analysis of polysomes
225(6)
References
231


Biežāk uzdotie jautājumi par e-grāmatām Biežāk uzdotie jautājumi par e-grāmatām
Permanent link: https://www.kriso.lv/db/97800808869612e.html
Keywords: