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PDF电子书：RNA Purification
目录：4 I% e7 ]/ k3 ?0 R" ?
Selecting a Purification Strategy . . . . . . . . . . . . . . . .. . . . . . . . . 1980 S. O4 Q; O. [
Do Your Experiments Require Total RNA or mRNA? . . . . . 198# p# [* E8 Z/ m+ D
Is It Possible to Predict the Total RNA Yield from
' R0 y0 s8 U# t8 l( pa Certain Mass of Tissue or Number of Cells? . . . . . . . . 201
& B$ H3 ^' g# }Is There Protein in Your RNA Preparation, and
1 K6 g$ P1 ]9 ]' x, r6 s+ ~: WIf So, Should You Be Concerned? . . . . . . . . . . . . . . . . . . . . 202
7 O8 q! B/ O& MIs Your RNA Physically Intact? Does It Matter? . . . . . . . . . . 2021 ~" w, i, D6 n0 {1 g2 G! R7 o
Which Total RNA Isolation Technique Is Most
4 F) b8 U/ M, ]! H( `Appropriate for Your Research? . . . . . . . . . . . . . . . . . . . . . 203
0 t: h+ x# ^- N* `. Y: T+ VWhat Protocol Modifications Should Be Used for
% z- A, ~$ _# o5 uRNA Isolation from Difficult Tissues? . . . . . . . . . . . . . . . . 2072 e5 l. s" s9 j4 ]3 @
Is a One-Step or Two-Step mRNA-(poly(A) RNA)-
7 B, W9 ?0 U8 F* h! Y# \Purification Strategy Most Appropriate for Your' m* W( b' g$ p
Situation? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2092 f& T: U2 k: k' k
How Many Rounds of Oligo(dT)–Cellulose- H* c# g" D% b: N' a; w
Purification Are Required? . . . . . . . . . . . . . . . . . . . . . . . . . 2107 i& s" w& C- C+ f! K
Which Oligo(dT)–Cellulose Format Is Most% O9 i# q1 c% {$ F& \" Q( I
Appropriate? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210
' x' c5 A3 Z9 Y' ?2 `Can Oligo(dT)–Cellulose Be Regenerated and Reused? . . . 211
) ^" I+ c5 G2 v: K- y M6 _0 a6 ~% FCan a Kit Designed to Isolate mRNA Directly from
' U7 ?0 D: E- E0 k; r" f; qthe Biological Sample Purify mRNA from Total RNA? . . . 212
# ?! H' x; u' i4 P9 K) s1 @Maximizing the Yield and Quality of an RNA Preparation . . . 212
- X2 F6 K- y% Y) b2 F2 a8 ]What Constitutes “RNase-Free Technique”? . . . . . . . . . . . . 212& U8 L. D) d, q; d( y5 b
How Does DEPC Inhibit RNase? . . . . . . . . . . . . . . . . . . . . . . 213+ E5 v" R+ l; e3 @8 H
How Are DEPC-Treated Solutions Prepared? Is$ o- _# {# V* T- Z
More DEPC Better? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2130 L& K8 I4 ]6 [! }8 n* n0 k' O. j& d" _
Should You Prepare Reagents with DEPC-Treated Water,4 m% P, G4 v2 b" x1 A, z
or Should You Treat Your Pre-made Reagents with: [* J5 A4 V4 T+ g9 v' @: ^5 d9 P
DEPC? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214) l( m9 X) P J. `' E
How Do You Minimize RNA Degradation during Sample) P4 F, v7 C4 I& s% \" @
Collection and Storage? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2145 j7 n! i3 |2 P9 Q; y" R
How Do You Minimize RNA Degradation during Sample6 y1 [# a% ~( z
Disruption? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 215; R+ K4 M+ G, ^ e: h( T3 V
Is There a Safe Place to Pause during an RNA
1 S8 z+ T; a8 {4 rPurification Procedure? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218
$ U% O; _0 j5 {3 d( x* g% BWhat Are the Options to Quantitate Dilute RNA! |4 _& O. M$ R# }. n2 j
Solutions? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218
7 i' E" S) d9 t) oWhat Are the Options for Storage of Purified RNA? . . . . . . . 219
' u/ U( G7 F' L+ V3 e$ uTroubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2209 \7 y) |. `* E) J$ l
A Pellet of Precipitation RNA Is Not Seen at the End of, ]" D" i' u8 A7 Y. c, P) G
the RNA Purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220/ O2 P) V0 O' B, B0 r
A Pellet Was Generated, but the Spectrophotometer" a' ~* G8 v! Y
Reported a Lower Reading Than Expected, or Zero
8 |0 _" d) y; ?' V9 ]+ MAbsorbance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
$ }) B6 T- m( F1 i/ e' j0 XRNA Was Prepared in Large Quantity, but it Failed$ s# Q$ [& j0 u
in a Downstream Reaction: RT PCR is an/ g' n, }' G% ?1 Y
Example . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
m! j+ [! b, K% o4 HMy Total RNA Appeared as a Smear in an Ethidum, o* b. P1 j. N# p% }5 r' {
Bromide-stained Denaturing Agarose Gel; 18S and
: E) H& Q+ u8 C+ Z* T28S RNA Bands Were not Observed . . . . . . . . . . . . . . . . 2224 E, O& z. C* ~ u2 e1 Q
Only a Fraction of the Original RNA Stored at -70°C
0 _4 ~: p& c$ e) eRemained after Storage for Six Months . . . . . . . . . . . . . . 222
8 k* [' a: C. G7 f3 \Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222 |
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发表于 2011-10-25 19:26
发表于 2011-10-26 08:59
