|
- 积分
- 19
- 威望
- 19
- 包包
- 934
|
PDF电子书：RNA Purification
目录：
5 o$ M" m/ w- o* F+ o9 YSelecting a Purification Strategy . . . . . . . . . . . . . . . .. . . . . . . . . 1987 p. X7 R% S$ `; n
Do Your Experiments Require Total RNA or mRNA? . . . . . 198' N* g2 O2 Z4 c1 b% w6 K
Is It Possible to Predict the Total RNA Yield from
6 m1 ?0 p" Q3 d! l6 Z" N( y1 Ta Certain Mass of Tissue or Number of Cells? . . . . . . . . 201 D2 H2 `! @2 k, S/ n) `8 ~
Is There Protein in Your RNA Preparation, and
# i" R$ Q- R# \, N1 sIf So, Should You Be Concerned? . . . . . . . . . . . . . . . . . . . . 202
, ]/ Z4 m! _. Z" R% c% v; v& CIs Your RNA Physically Intact? Does It Matter? . . . . . . . . . . 202
1 k" n- |; \$ G4 o7 G WWhich Total RNA Isolation Technique Is Most7 |' `6 Y" f+ {9 V: `5 K/ |3 d
Appropriate for Your Research? . . . . . . . . . . . . . . . . . . . . . 203
! w: [" j, |8 ^What Protocol Modifications Should Be Used for) r5 B o1 z* i/ q ^' e
RNA Isolation from Difficult Tissues? . . . . . . . . . . . . . . . . 207
. N" X/ e( I% K8 e' z3 n$ wIs a One-Step or Two-Step mRNA-(poly(A) RNA)-
* ]4 d/ M1 q$ F2 Z+ VPurification Strategy Most Appropriate for Your( S2 w7 P" E0 t+ i
Situation? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209; c0 z) X7 g/ B; I2 F
How Many Rounds of Oligo(dT)–Cellulose! J" P- Z& ?. s7 ~
Purification Are Required? . . . . . . . . . . . . . . . . . . . . . . . . . 210) F* j, H7 K- s" c! x, v
Which Oligo(dT)–Cellulose Format Is Most" b, ?/ Y: j4 ^
Appropriate? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210* U, |# o0 l: [5 v9 {% j, k
Can Oligo(dT)–Cellulose Be Regenerated and Reused? . . . 211) @7 B: K) b) ]+ W
Can a Kit Designed to Isolate mRNA Directly from
6 t J. a# R( {, rthe Biological Sample Purify mRNA from Total RNA? . . . 212
6 W9 n2 c3 `- X/ i3 }9 iMaximizing the Yield and Quality of an RNA Preparation . . . 2126 i- P. L# V: _' p' g, u* t/ O
What Constitutes “RNase-Free Technique”? . . . . . . . . . . . . 212
, Q/ z8 c- |$ zHow Does DEPC Inhibit RNase? . . . . . . . . . . . . . . . . . . . . . . 213
" |! M% M, Y! o" |- x iHow Are DEPC-Treated Solutions Prepared? Is
( L8 Q+ l- _0 _3 k. n SMore DEPC Better? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213% }4 A* P9 x6 r' w6 a9 o, K$ b
Should You Prepare Reagents with DEPC-Treated Water,
. F0 e1 X! J1 [1 Z: zor Should You Treat Your Pre-made Reagents with3 k; T3 Y# A5 |- ^( R/ o- F2 x$ w
DEPC? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2140 l8 l% k. Z6 U
How Do You Minimize RNA Degradation during Sample
1 J; w' N7 R. L1 p \Collection and Storage? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214
u) v4 w( p8 g! g3 u3 FHow Do You Minimize RNA Degradation during Sample
" M4 A: z' W4 PDisruption? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 215
& [/ V+ ~ \6 r# {8 b0 l q& KIs There a Safe Place to Pause during an RNA: M9 l: n5 ~8 E7 m! X6 N0 W' I
Purification Procedure? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218
) k- }2 r0 `& X$ p/ aWhat Are the Options to Quantitate Dilute RNA B0 N9 T; z% s6 w9 V0 k" D
Solutions? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218# K0 {- A4 y4 G: s! O& H
What Are the Options for Storage of Purified RNA? . . . . . . . 2193 O% r7 K; C0 E1 H# I( _2 e8 i% K5 r
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
; M" ]7 i" q# Q* |% O+ J" RA Pellet of Precipitation RNA Is Not Seen at the End of( s; D8 u U& F
the RNA Purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220+ p$ [( P) }! F, Q2 |
A Pellet Was Generated, but the Spectrophotometer6 Q' B3 ~ _% ^7 ?: P0 n
Reported a Lower Reading Than Expected, or Zero
5 Q' J# b% W, s; y/ B g1 h' oAbsorbance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2213 N( Q( @4 `9 n
RNA Was Prepared in Large Quantity, but it Failed ~3 y" {4 |5 ^+ f
in a Downstream Reaction: RT PCR is an) U# ?1 W" Z/ G
Example . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
$ E5 a* I! w1 ^$ _) zMy Total RNA Appeared as a Smear in an Ethidum
) Y" m3 _7 @- H S: |% |Bromide-stained Denaturing Agarose Gel; 18S and
' O# _. K' w9 f, g, _5 ?28S RNA Bands Were not Observed . . . . . . . . . . . . . . . . 2228 `& {& x. [4 N& d1 T& h D6 ~
Only a Fraction of the Original RNA Stored at -70°C
+ `7 c( f) T2 YRemained after Storage for Six Months . . . . . . . . . . . . . . 2220 v* _! E3 }) P4 E1 E( |- C0 B, q
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222 |
|
发表于 2011-10-25 19:26
发表于 2011-10-26 08:59
