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PDF电子书：RNA Purification
目录：6 V, [( v; c8 e6 z4 |
Selecting a Purification Strategy . . . . . . . . . . . . . . . .. . . . . . . . . 198
9 M( L! d* C: U7 w g) ZDo Your Experiments Require Total RNA or mRNA? . . . . . 198
# H4 D' m1 o+ \3 t) k8 C AIs It Possible to Predict the Total RNA Yield from+ E. V+ V1 x/ M
a Certain Mass of Tissue or Number of Cells? . . . . . . . . 2019 n2 W. \9 `- ?4 ?; c) }
Is There Protein in Your RNA Preparation, and3 w+ C9 j9 T, Y* [/ s. h' e+ x1 s
If So, Should You Be Concerned? . . . . . . . . . . . . . . . . . . . . 2026 ^2 H. k' A0 R, m% p3 M8 {0 e
Is Your RNA Physically Intact? Does It Matter? . . . . . . . . . . 202. h1 B8 ^2 |5 Q* ~$ y" G5 A) f
Which Total RNA Isolation Technique Is Most, F" c/ i7 L+ R! S& g2 s
Appropriate for Your Research? . . . . . . . . . . . . . . . . . . . . . 203
Z k+ ~% a% xWhat Protocol Modifications Should Be Used for8 D" e. B4 i0 D8 C2 C
RNA Isolation from Difficult Tissues? . . . . . . . . . . . . . . . . 207
: `: ?1 D1 x! C( O( DIs a One-Step or Two-Step mRNA-(poly(A) RNA)-
7 B4 a, ~+ h2 XPurification Strategy Most Appropriate for Your
4 I& U* N, k9 b% YSituation? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2095 N, b _: O& U* J
How Many Rounds of Oligo(dT)–Cellulose
; l4 x$ t- d) w" z( DPurification Are Required? . . . . . . . . . . . . . . . . . . . . . . . . . 210
1 b& H" a, N. I, r) DWhich Oligo(dT)–Cellulose Format Is Most4 U* R, a. x: ?0 _% w
Appropriate? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2104 o( B' R4 J) ^9 {7 \
Can Oligo(dT)–Cellulose Be Regenerated and Reused? . . . 211% f+ M( ~" B" S) }2 I
Can a Kit Designed to Isolate mRNA Directly from
# s- o- Q2 n$ z- S& ^8 Fthe Biological Sample Purify mRNA from Total RNA? . . . 212* c( A' k m; _' I+ @6 z
Maximizing the Yield and Quality of an RNA Preparation . . . 212
% H6 h# v9 x8 ]7 D FWhat Constitutes “RNase-Free Technique”? . . . . . . . . . . . . 2128 g0 N$ s# p: [0 t. K( u
How Does DEPC Inhibit RNase? . . . . . . . . . . . . . . . . . . . . . . 213' h6 \+ ^$ `8 B7 u. T& [6 Q3 F7 M' c7 T
How Are DEPC-Treated Solutions Prepared? Is
. W) ~4 X: R5 @6 AMore DEPC Better? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
) F+ z, Z8 C4 }/ XShould You Prepare Reagents with DEPC-Treated Water,
1 f0 Z: B, Q0 h/ ~" b7 B, Tor Should You Treat Your Pre-made Reagents with
( K2 `9 U( }) ]DEPC? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214
3 d5 f# z1 d: K! R1 uHow Do You Minimize RNA Degradation during Sample
% ~+ z" d+ W& SCollection and Storage? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214* _4 t4 D: S% N' u5 `! T6 H
How Do You Minimize RNA Degradation during Sample: Q" G2 F- ]5 O0 S4 k
Disruption? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2158 v: c$ Q, i0 S# i/ p
Is There a Safe Place to Pause during an RNA
5 X: \' {8 j5 [" _9 ^$ oPurification Procedure? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2185 `4 S A5 O/ U- N
What Are the Options to Quantitate Dilute RNA
, s% \) N2 z( V- X3 G% NSolutions? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218- w' L. q4 x: L; M; _- I
What Are the Options for Storage of Purified RNA? . . . . . . . 219 N* Q; v9 h3 s! g
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2200 ^5 ^' z* W% r% O* F$ R" v
A Pellet of Precipitation RNA Is Not Seen at the End of' [. W6 l+ b' g
the RNA Purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
6 i, U$ b* O5 g+ RA Pellet Was Generated, but the Spectrophotometer, a! Y! c; Z- ?0 Z+ b
Reported a Lower Reading Than Expected, or Zero
- i) J& n, G8 ^& L- v4 f7 bAbsorbance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
6 \- |* }5 T; \9 l6 rRNA Was Prepared in Large Quantity, but it Failed- ]# F0 S7 Y% i2 E' l& ?
in a Downstream Reaction: RT PCR is an
2 s% \- [. J, ~5 w% IExample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2219 }( H+ V4 h& |+ `% T8 z- q
My Total RNA Appeared as a Smear in an Ethidum' ~0 w& ~8 k" ?/ W2 b7 ?
Bromide-stained Denaturing Agarose Gel; 18S and# T! [7 \8 z& j/ V
28S RNA Bands Were not Observed . . . . . . . . . . . . . . . . 222
* E$ @' b6 B0 _Only a Fraction of the Original RNA Stored at -70°C$ _. p6 ~2 c) w" T) E
Remained after Storage for Six Months . . . . . . . . . . . . . . 222: g% o) k2 o0 t- E8 x5 z4 d5 E
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222 |
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发表于 2011-10-25 19:26
发表于 2011-10-26 08:59
