Biocompare Console Feed

Transfection Reagents

Sun, 05 Oct 2008 12:51:00 GMT

Finding the right transfection reagent is a key step in your cell-based expression studies. You need a reagent that will effectively deliver your construct without causing cellular changes that could interfere with the interpretation of your results. In addition to these essentials, it'd be nice...

Sample Management

Tue, 30 Sep 2008 16:24:00 GMT

Sample management software has become a must-have for large and small laboratories. When you are talking about precious resources and valuable person hours, it is no longer acceptable to keep track of samples by memory, or by a piece of paper scotch-taped...

Options for MS Fragmentation

Thu, 25 Sep 2008 11:25:00 GMT

Try this thought exercise: Imagine you're blindfolded, and asked to identify the object someone puts into your hand. The object is a bottle filled with jellybeans. Given its texture, weight, and shape, you probably can surmise you've been handed a bottle. But what are its contents? To figure that out...

Choosing The Best Genomic DNA Purification Kit

Wed, 01 Oct 2008 15:54:00 GMT

These days, a broad range of DNA purification kits are available from various vendors which makes the selection of the best DNA purification kit a daunting task. However, on a positive note, it gives us more opportunity to select the most appropriate kit to meet the challenges posed by sample sources and volumes, throughput levels and downstream applications. When selecting a DNA purification kit, there are several factors to keep in mind. The first thing to consider is the source of genomic DNA. Some kits are able to extract DNA from a wide variety of samples such as blood, tissue, plant or bacterial cells. However, others are designed for a specific tissue type or even for a specific species. Secondly, what is the starting volume of the sample and expected DNA yield? Kits are available in micro, mini, midi and maxi formats; each has a limitation on the volume of starting material and amount of DNA eluted. Thirdly, you want to be sure that the purity of eluted DNA is suitable for your application; for example, some protocols use organic solvent(s) and traces of which can remain in eluted DNA and possibly interfere with sensitive downstream applications. Other factors take into consideration are how many samples you wish to process at a time and frequency of kit use as some kits may be more suitable for managing multiple samples at once and are also available in high-throughput or automated formats. You might also want to compare the time and cost effectiveness of the kit. Overall, I recommend you go through the manual and protocol of the kit and compare the facts mentioned above. Also, don't hesitate to contact the company for additional information. Below is a comparison of a few DNA purification kits to get you started.

Anupriya Agarwal
Postdoctoral Fellow
Oregon Health and Science University

RT-PCR Kits

Wed, 01 Oct 2008 15:57:00 GMT

Reverse transcription is the process of turning RNA molecules into complementary DNA (cDNA). One major advantage of using cDNA over RNA in the study of gene expression is that cDNA is more stable than RNA. When choosing an RT-PCR kit, you want your money to turn into reliable results, especially in quantitative experiments. The first rule for RT-PCR is to use high quality starting material (RNA). Therefore, choose a kit that fits your specific need: long templates, viral RNA, quantitative assays, etc. Different kits may require different starting amount (may vary from 1 ug to 5 ug) of RNA. I have obtained abundant cDNA for most subsequent applications when using both 1 ug and 5 ug of starting material, provided that the kit was properly stored. Another issue you may need to notice is whether the kit is 'one-step' or 'two-step'. For 'two-step' kits, you will first use reverse transcriptase to generate cDNA from total RNA or mRNA, and then you can perform gene-specific PCR in separate reactions. For 'one-step' kits, the enzyme blend includes both thermostable reverse transcriptase and DNA polymerase, so that you can use gene-specific primers instead of random hexamer or oligo-dT; this allows faster detection of the target gene. Finally, some kits provide RNase inhibitor during the RT reaction, and RNase for degradation of the RNA molecules after the reaction. Inclusion of such reagents is icing on the cake. Finally, once you have chosen a kit, stick to it until the end of your experiment. Then everything is going to be fine.

Issan Tam
Research Assistant
The Chinese University of Hong Kong

5'/3' RACE Kit From Roche Applied Science

Wed, 01 Oct 2008 15:15:00 GMT

Recently, I investigated the possibility of a novel fusion protein in lung tumors. Since I had sequence information on one fusion partner, I was able to verify the presence of the 3' end of its transcript with RT-PCR. I then wanted to know...

Comments (0) - Read and Add your own Comments