【资源】RNA Purification

Do Your Experiments Require Total RNA or mRNA?
看看就会有收获
Selecting a Purification Strategy . . . . . . . . . . . . . . . .. . . . . . . . . 198
Do Your Experiments Require Total RNA or mRNA? . . . . . 198
Is It Possible to Predict the Total RNA Yield from
a Certain Mass of Tissue or Number of Cells? . . . . . . . . 201
Is There Protein in Your RNA Preparation, and
If So, Should You Be Concerned? . . . . . . . . . . . . . . . . . . . . 202
Is Your RNA Physically Intact? Does It Matter? . . . . . . . . . . 202
Which Total RNA Isolation Technique Is Most
Appropriate for Your Research? . . . . . . . . . . . . . . . . . . . . . 203
What Protocol Modifications Should Be Used for
RNA Isolation from Difficult Tissues? . . . . . . . . . . . . . . . . 207
Is a One-Step or Two-Step mRNA-(poly(A) RNA)-
Purification Strategy Most Appropriate for Your
Situation? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209
How Many Rounds of Oligo(dT)–Cellulose
Purification Are Required? . . . . . . . . . . . . . . . . . . . . . . . . . 210
Which Oligo(dT)–Cellulose Format Is Most
Appropriate? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210
Can Oligo(dT)–Cellulose Be Regenerated and Reused? . . . 211
Can a Kit Designed to Isolate mRNA Directly from
the Biological Sample Purify mRNA from Total RNA? . . . 212
Maximizing the Yield and Quality of an RNA Preparation . . . 212
What Constitutes “RNase-Free Technique”? . . . . . . . . . . . . 212
How Does DEPC Inhibit RNase? . . . . . . . . . . . . . . . . . . . . . . 213
How Are DEPC-Treated Solutions Prepared? Is
More DEPC Better? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
Should You Prepare Reagents with DEPC-Treated Water,
or Should You Treat Your Pre-made Reagents with
DEPC? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214
How Do You Minimize RNA Degradation during Sample
Collection and Storage? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214
How Do You Minimize RNA Degradation during Sample
Disruption? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 215
Is There a Safe Place to Pause during an RNA
Purification Procedure? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218
What Are the Options to Quantitate Dilute RNA
Solutions? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218
What Are the Options for Storage of Purified RNA? . . . . . . . 219
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
A Pellet of Precipitation RNA Is Not Seen at the End of
the RNA Purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
A Pellet Was Generated, but the Spectrophotometer
Reported a Lower Reading Than Expected, or Zero
Absorbance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
RNA Was Prepared in Large Quantity, but it Failed
in a Downstream Reaction: RT PCR is an
Example . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
My Total RNA Appeared as a Smear in an Ethidum
Bromide-stained Denaturing Agarose Gel; 18S and
28S RNA Bands Were not Observed . . . . . . . . . . . . . . . . 222
Only a Fraction of the Original RNA Stored at -70°C
Remained after Storage for Six Months . . . . . . . . . . . . . . 222
Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222