Aging Aging

More documents

Recommendations

Info

Mitochondrial DNA Mutations 253Fig. 2. Primer-shift PCR from single muscle fibers showing the combination ofPCR primers (pairs a–h) used to screen the mtDNA genome. A shows the productsgenerated from total DNA isolated from skeletal muscle of a patient with multiplemtDNA deletions. B and C represent the products generated from the DNA isolatedfrom two individual COX-deficient muscle fibers, highlighting the presence of differentdeleted mtDNA species in each fiber.L8197 (nt 8197–8216), H466 (nt 466–447), H9050 (nt 9050–9031), H13640 (nt13640–13621), H14840 (nt 14840–14821), H15260 (nt 15260–15241), H16150(nt 16150–16131), and H16560 (nt 16560–16541). Stock solutions (20 µM) arestored at –20°C.
254 Taylor et al.4. Sterile water.5. Ice: All reactions are set up on ice.6. Horizontal gel electrophoresis equipment.7. 1.5% Agarose gel containing ethidium bromide; 1× TAE running buffer.3. Methods3.1. Long-Range PCR of mtDNA1. Thaw all components stored at –20°C and bring reaction buffer 3 to room temperature.Vortex well to mix, checking the reaction buffer for crystals. If crystalsare present leave at room temperature overnight and mix well again. Place all thecomponents on ice.2. Label two, sterile 1.5-mL Eppendorf tubes A and B; the master mix is made upas two separate mixes to avoid degradation of the primers or template due to the3'→5' exonuclease activity of the Pwo enzyme.3. Combine the following components to form mix A for each 50 µL PCR:2.5 µL 10 mM dATP2.5 µL 10 mM dCTP2.5 µL 10 mM dGTP2.5 µL 10 mM dTTP1.5 µL 10µM Forward primer1.5 µL 10µM Reverse primer12 µL Sterile water4. Vortex-mix briefly, centrifuge the mix, and aliquot 25 µL into each thin-walledtube on ice.5. Add 1 µL of DNA template to each separate tube.6. Prepare master mix B; for each 50-µL reaction combine the following:5 µL 10× Reaction buffer 310 µL 1 mg/mL of BSA8.65 µL Sterile water*0.35 µL Expand enzyme*While the kit recommends using 0.7 µL of enzyme per 50 µL of PCR reaction,we find that the same efficiency of amplification is achieved using only 0.35 µL.Vortex-mix and briefly centrifuge the mix, aliquoting 24 µL into each thinwalledtube, mixing gently to avoid introducing air bubbles.7. If necessary overlay the reactions with 30 µL of mineral oil.8. Centrifuge the samples briefly, and place back on ice.9. Begin the following PCR program to heat the block to 92–94°C:Initial denaturation 92–94°C* 3 min10 Cycles 92–94°C* 10–30 s55–68°C** 30 s
Page 7:
6 Strehlerlead to possible death si
Page 10 and 11:
Understanding Aging 9the intestine
Page 12 and 13:
Understanding Aging 117. Do long-li
Page 14 and 15:
Understanding Aging 13is, the great
Page 16 and 17:
Understanding Aging 15While this re
Page 18 and 19:
Understanding Aging 172. Pearl, R.
Page 20 and 21:
Understanding Aging 1945. Perovic,
Page 22 and 23:
24 Cristofalo, Volker, and Allenmen
Page 24 and 25:
26 Cristofalo, Volker, and Allenrec
Page 26 and 27:
28 Cristofalo, Volker, and Allenski
Page 28 and 29:
30 Cristofalo, Volker, and AllenTab
Page 30 and 31:
Page 32 and 33:
34 Cristofalo, Volker, and AllenNa
Page 34 and 35:
Page 36 and 37:
38 Cristofalo, Volker, and Allentak
Page 38 and 39:
40 Cristofalo, Volker, and Allenb.
Page 40 and 41:
42 Cristofalo, Volker, and Allenand
Page 42 and 43:
44 Cristofalo, Volker, and AllenRef
Page 44 and 45:
46 Cristofalo, Volker, and Allen33.
Page 46 and 47:
48 Cristofalo, Volker, and Allen61.
Page 48 and 49:
50 Cristofalo, Volker, and Allenhum
Page 50 and 51:
52 Cristofalo, Volker, and Allen125
Page 52 and 53:
54 Pawelecdiluted cells on an irrad
Page 54 and 55:
56 Pawelec3. Method3.1. Source of C
Page 56 and 57:
58 Pawelecweekly or fortnightly sub
Page 58 and 59:
60 Pawelection, the VERUM Foundatio
Page 60 and 61:
Telomeres and Replicative Senescenc
Page 62 and 63:
Page 64 and 65:
Page 66 and 67:
Page 68 and 69:
Detection of Molecular Events 715De
Page 70 and 71:
Detection of Molecular Events 73hav
Page 72 and 73:
Detection of Molecular Events 75Fig
Page 74 and 75:
Detection of Molecular Events 772.
Page 76 and 77:
Detection of Molecular Events 79Fig
Page 78 and 79:
Page 80 and 81:
Page 82 and 83:
86 Kirk and Millerexperiments on un
Page 84 and 85:
88 Kirk and MillerFig. 1. (A) Analy
Page 86 and 87:
90 Kirk and MillerFurthermore, limi
Page 88 and 89:
92 Kirk and Millersufficiently dilu
Page 90 and 91:
94 Kirk and Miller4. Run samples on
Page 92 and 93:
96 Kirk and Miller16. Lange-Carter,
Page 94 and 95:
98 Engel, Adibzadeh, and PawelecPCR
Page 96 and 97:
100 Engel, Adibzadeh, and Pawelec5.
Page 98 and 99:
102 Engel, Adibzadeh, and PawelecTa
Page 100 and 101:
104 Engel, Adibzadeh, and PawelecTa
Page 102 and 103:
106 Engel, Adibzadeh, and Pawelecam
Page 104 and 105:
Page 106 and 107:
110 Engel, Adibzadeh, and Pawelecfo
Page 108 and 109:
Page 110 and 111:
114 Engel, Adibzadeh, and Pawelecha
Page 112 and 113:
Xenobiotic-Metabolizing Enzymes 119
Page 114 and 115:
Page 116 and 117:
Page 118 and 119:
Page 120 and 121:
Page 122 and 123:
Page 124 and 125:
Assessing Antioxidant Status 1339As
Page 126 and 127:
Assessing Antioxidant Status 135Fig
Page 128 and 129:
Assessing Antioxidant Status 137Tab
Page 130 and 131:
Assessing Antioxidant Status 139add
Page 132 and 133:
Assessing Antioxidant Status 14121.
Page 134 and 135:
Comet Assay of DNA Damage 14310Meas
Page 136 and 137:
Comet Assay of DNA Damage 145contro
Page 138 and 139:
Comet Assay of DNA Damage 1476. Lys
Page 140 and 141:
Comet Assay of DNA Damage 1494. Gen
Page 142 and 143:
Comet Assay of DNA Damage 151anode
Page 144 and 145:
Comet Assay of DNA Damage 1535. As
Page 146 and 147:
Comet Assay of DNA Damage 15515. In
Page 148 and 149:
Comet Assay of DNA Damage 15720. Co
Page 150 and 151:
160 van der Schanswithin 1 h, after
Page 152 and 153:
162 van der Schans10. High-binding
Page 154 and 155:
164 van der Schans3. Neutralize aft
Page 156 and 157:
166 van der SchansThe calculations
Page 158 and 159:
8-oxoguanine Levels in Nuclear DNA
Page 160 and 161:
Page 162 and 163:
Page 164 and 165:
Page 166 and 167:
Mutation and the Aging Process 1791
Page 168 and 169:
Mutation and the Aging Process 181m
Page 170 and 171:
Mutation and the Aging Process 183b
Page 172 and 173:
Page 174 and 175:
Page 176 and 177:
190 HouThe HPRT mutational spectrum
Page 178 and 179:
192 Houamplifications are carried o
Page 180 and 181:
194 Hou4. Reading: Load 10 µL of t
Page 182 and 183:
196 Hou4.2. MP-PCRUse preferably DN
Page 184 and 185: Susceptibility of LDL to Oxidation
Page 194 and 195: Analysis of Pentosidine 20916Measur
Page 196 and 197: Analysis of Pentosidine 211Fig. 2.
Page 198 and 199: Analysis of Pentosidine 213Fig. 4.
Page 200 and 201: Analysis of Pentosidine 2152. Preci
Page 202 and 203: Analysis of Pentosidine 2177. Nagar
Page 204 and 205: 222 Miquellipoperoxides and malonal
Page 206 and 207: 224 Miquelat 4°C. Protect this sol
Page 208 and 209: 226 Miquel3.1. Animal AnesthesiaMed
Page 210 and 211: 228 Miquelsteps to carry out a conv
Page 212 and 213: 230 Miquelsuspending it over the so
Page 214 and 215: 232 Miquelcontrast, washing with bu
Page 216 and 217: 234 MiquelFig. 3. Electron microsco
Page 218 and 219: Damage to Mitochondria 23718Causes
Page 220 and 221: Damage to Mitochondria 239Fig. 1. E
Page 222 and 223: Damage to Mitochondria 2412. To obt
Page 224 and 225: Damage to Mitochondria 243synthesis
Page 226 and 227: Mitochondrial DNA Mutations 24519An
Page 228 and 229: Mitochondrial DNA Mutations 247F.P.
Page 230 and 231: Mitochondrial DNA Mutations 249desi
Page 232 and 233: Mitochondrial DNA Mutations 2514. M
Page 236 and 237: Mitochondrial DNA Mutations 25568°
Page 238 and 239: Mitochondrial DNA Mutations 257Fig.
Page 240 and 241: Mitochondrial DNA Mutations 2593. P
Page 242 and 243: Mitochondrial DNA Mutations 261X-10
Page 244 and 245: Mitochondrial DNA Mutations 26315.
Page 246 and 247: Mitochondrial DNA Mutations 26520An
Page 248 and 249: Mitochondrial DNA Mutations 267Tabl
Page 250 and 251: Mitochondrial DNA Mutations 269muta
Page 252 and 253: Mitochondrial DNA Mutations 2717. P
Page 254 and 255: Mitochondrial DNA Mutations 2733.2.
Page 256 and 257: Mitochondrial DNA Mutations 275that
Page 258 and 259: Mitochondrial DNA Mutations 27711.
Page 260 and 261: 282 BeckmanWhatever the mechanism e
Page 262 and 263: 284 BeckmanT-cell adherence to vasc
Page 264 and 265: 286 BeckmanT3 hybridoma cells satis
Page 266 and 267: 288 BeckmanHaving identified a cand
Page 268 and 269: 290 Beckmanthe plates vigorously wi
Page 270 and 271: 292 Grubeck-Loebenstein, Saurwein-T
Page 284 and 285:
306 Grubeck-Loebenstein, Saurwein-T
Page 286 and 287:
308 Grubeck-Loebenstein, Saurwein-T
Page 288 and 289:
NK Cell Function in Aging 31123Age-
Page 290 and 291:
NK Cell Function in Aging 3135. Pet
Page 292 and 293:
NK Cell Function in Aging 31518. Io
Page 294 and 295:
NK Cell Function in Aging 3173.3. C
Page 296 and 297:
NK Cell Function in Aging 3199mL of
Page 298 and 299:
Immunogenetics and Life-Span 32124I
Page 300 and 301:
Immunogenetics and Life-Span 323Tab
Page 302 and 303:
Immunogenetics and Life-Span 325rea
Page 304 and 305:
Immunogenetics and Life-Span 3273.
Page 306 and 307:
Page 308 and 309:
Immunogenetics and Life-Span 3313.5
Page 310 and 311:
Page 312 and 313:
Page 314 and 315:
Page 316 and 317:
Page 318 and 319:
Page 320 and 321:
Page 322 and 323:
Page 324 and 325:
Page 326 and 327:
Immunogenetics and Life-Span 349to
Page 328 and 329:
354 Yuto-implement intervention and
Page 330 and 331:
356 Yuof their quality control proc
Page 332 and 333:
358 Yugenesis of major diseases (1-
Page 334 and 335:
Genetically Engineered Mice 36126Th
Page 336 and 337:
Genetically Engineered Mice 363as t
Page 338 and 339:
Genetically Engineered Mice 365in b
Page 340 and 341:
Genetically Engineered Mice 367numb
Page 342 and 343:
Genetically Engineered Mice 369mula
Page 344 and 345:
Genetically Engineered Mice 371Othe
Page 346 and 347:
Genetically Engineered Mice 373locu
Page 348 and 349:
Genetically Engineered Mice 37537.
Page 350:
Genetically Engineered Mice 377→
show all

Aging Aging

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?