Molecular Biology Problem Solver 11 pps

... conditions, as source of problems, 21–22, 40–42 Storage phosphor imagers dynamic range of, 442–443 erasure of, 447 in nucleic acid hybridization, 441–448 operation of, 441 problems with, 447–448 quantitative ... 350–353 native PAGE versus, 345–348 problems with, 480–481 standardized gels for, 363 Sealed containment rooms, in biosafety, 117 Secondary antibodies, 384 problems with, 395 Secondar...

Ngày tải lên: 02/07/2014, 04:21

... pH of 7 and 10 would be acceptable. Analyz- ing samples covering a wider range will be more accurately measured with a multiple-point calibration covering a range such as 4, 7, and 10, or even ... 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

... . . . . . . . . . . 150 A Wipe Test Detected Radioactivity on the Outside of the Vial. Does This Indicate a Problem? . . . . . . . . . . . . 151 Molecular Biology Problem Solver: A Laboratory ... Requirements of Autoclaving? The minimum requirements for sterilization are 15 lb of pres- sure per square inch at 115 C for 15 minutes of sterilization time (not including the exhaus...

Ngày tải lên: 02/07/2014, 04:21

... . . . . . . . 164 How Can You Concentrate a Radioactive Solution? . . . . . . 164 Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166 Appendix ... . . . . . . . . . . . . 161 What Can You Do to Achieve Minimum Radioactive Dose? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162 How Can You Organ...

Ngày tải lên: 02/07/2014, 04:21

... . . . . . . 185 Silica Resin-Based Strategies . . . . . . . . . . . . . . . . . . . . . . . . . . . 186 Isolation from Electrophoresis Gels . . . . . . . . . . . . . . . . . . . . 187 What Are ... 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 extr...

Ngày tải lên: 02/07/2014, 04:21

... 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

... 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 discuss the ... RNA- binding filters. Increasing the ratio of sample mass: volume of lysis buffer can help alleviate this problem in filter-based isolations. Multiple acid–phenol extractions can be done to ensure...

Ngày tải lên: 02/07/2014, 04:21

... 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 resolving power. Suppose ... primer degradation by the 3¢–5¢ exonuclease activity (de Noronha and Mullins, 1992 and Skerra, 1992).The problem of reduced yield can be corrected by including an enzyme w...

Ngày tải lên: 02/07/2014, 04:21

... way to store 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–thawing ... sample in smaller quantity lessens the effect whereas the effect worsens with more sample. If the problem is caused by template modification, dilution will have no effect. Compounds such as N-ph...

Ngày tải lên: 02/07/2014, 04:21

... considering protein blot quantitation. Standards (known amounts of purified target protein—not to be confused with molecular weight stan- dards) must be run on every blot, since even with the most con- sistent technique ... accurate records. Consistency and good record-keeping will make it much easier to isolate the source of any problem that may come up later. Which Transfer Membrane Is M...

Ngày tải lên: 02/07/2014, 04:21

... Problems are inevitable, so taking a systematic approach to troubleshooting will, in the long run, save time, Western Blotting 389 Western Blotting 391 Protein remains in the gel Were there problems ... problems with contact or arrangement of the blotting apparatus? Were there problems with reagents? Has the protein high molecular weight? Was transfer time sufficient? Are the buffer componen...

Ngày tải lên: 02/07/2014, 04:21

... under-represented when working with screens and flours. When working with radioactive labels, this problem can be cor- rected by a combination of exposure at -70°C and sensitizing the film with a ... Autoradiography Film Static electricity can produce background signals on film. A solution to this problems has been proposed by Register (1999). The use of fluorescent crayolas to mark the orienta...

Ngày tải lên: 02/07/2014, 04:21

... Molecular Biology Labora- tory. Meth. Mol. Cell. Biol. 23:317–356. Ausubel, M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., and Struhl, K. 1993. Current Protocols in Molecular Biology. ... the information presented in Table 14.2 are two: • Problems at any one or combination of steps can generate inadequate hybridization data. • Problems at different stages of a hybridizat...

Ngày tải lên: 02/07/2014, 04:21