Molecular Biology Problem Solver 20 pps

... purification procedures are reviewed in O’Kennedy et al. (200 0), Neudecker and Grimm (200 0), Monteiro et al. (1999), Ferreira et al., 200 0. Ferreira et al. (1999), Ferreira et al. (1998), Huber ... contaminant is at least 20 times smaller than the smallest molecule to be purified. • Desalting, where the size difference between ions and bio- molecule is >>1 :20, works well even at...

Ngày tải lên: 02/07/2014, 04:21

... yield from, 201 202 , 203 206 , 207 209 Titer assays, troubleshooting with, 255–259 Toluene, radioisotopes in, 147 Top-loader balances, 51 Total RNA, purification of, 198 201 , 203 206 , 207 209 , 212 Toxicity, ... of RNA from, 202 203 lysis in, 215 maximizing yield from, 212–219 pauses during, 218 predicting yield from, 201 202 storing RNA from, 219 strategies for, 198–212 troublesh...

Ngày tải lên: 02/07/2014, 04:21

... Equipment 87 change due to temperature as they would in a manual calibration. A second source of problems is not matching a buffer or group of buffers with the correct buffer set in the meter.

Ngày tải lên: 02/07/2014, 04:21

... causing the problem. Is the Sample the Problem? If the sample reading seems inappropriate, measure the pH of a buffer standard. A correct measurement of the standard points to a sample problem. ... recom- mended. Sometimes the “expected” value is not obtained for a mea- surement but the correct value is. The problem is simply an incorrect perceived value. Competing ions, sodium ions...

Ngày tải lên: 02/07/2014, 04:21

... the Vial. Does This Indicate a Problem? . . . . . . . . . . . . 151 Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S. Gerstein Copyright © 200 1 by Wiley-Liss, Inc. ISBNs: ... autoclave drains. Allow Sufficient Time A liquid cycle (20 minutes of sterilization time) usually requires approximately 35 minutes. A wrapped (dry with fast exhaust) cycle (20 minutes of...

Ngày tải lên: 02/07/2014, 04:21

... postreference. This number should be 0. 820. Multiply your starting radioactive concentration by this fraction to obtain the new radioactive concentration: 15 mCi/ml ¥ 0. 820 = 12.3mCi/ml The total amount ... fully below. A Wipe Test Detected Radioactivity on the Outside of the Vial. Does This Indicate a Problem? If you detect contamination on the outside of your vial, contact your RSO. S...

Ngày tải lên: 02/07/2014, 04:21

... DNA and subsequent ethanol washes eliminate such a contaminant. Phenol can also be problematic. If you experience problems with DNA purified by a phenol-based strategy, apply chloroform to extract ... will slow you down or cause an accident. If you can’t smoothly do the motion in 10 seconds, take 20. You’ll improve through time and all along will be well aware of your estimated dose. You’ll ....

Ngày tải lên: 02/07/2014, 04:21

... is severely compromised. For example, even a single cleavage in 20% of the target molecules will decrease the signal on a North- ern blot by 20% . Nuclease protection assays and RT-PCR analy- ses will ... total RNA isolation. In this section we address problems presented by difficult tissues and offer troubleshooting techniques to help over- come these problems. A separate section will discu...

Ngày tải lên: 02/07/2014, 04:21

... DNA Absent Present 150 at 96°C polymerase (Dynazyme TM ) e Platinum Pfx f Present Absent 720 at 95°C 100 200 100–300 180 at 100°C Platinum Taq f Absent Present 96 at 95°C 50–60 60–150 Advantaq ... electrophoresis may not allow you to reach the conclusion that there are two different mRNA forms if their molecular weights are similar.You might require a separation technique with greater reso...

Ngày tải lên: 02/07/2014, 04:21

... Matrix Primer Forward Primer concentration (nM) 100 120 140 160 180 200 100 + ++++++++ 120 + ++++++++ 140 + + ++ ++ ++ +++ 160 + ++++++ 180 - 200 - - - - of 200 nM and reverse primer at 140 nM. Both the ... plasmid DNA (or control cDNA or genomic DNA) is in TE with 20 mg/ml glycogen (molecular biology grade, nuclease free) in small aliquots in a -20 C freezer. Repeated freeze–thaw...

Ngày tải lên: 02/07/2014, 04:21