Navigation Links
Advantages of ReadyAgarose Precast Gels Over 96-Well Handcast Gels for High-Throughput Analysis, Rev A

Divya Sharma1 and Carolyn Seggerson2, 1Tech/Aid, 7700 Edgewater Drive, Oakland, CA 94621 USA, and 2Bio-Rad Laboratories, Inc., 2000 Alfred Nobel Drive, Hercules, CA 94547 USA

Running PCR samples on agarose gels is a routine yet timeconsuming procedure. It is common to hand cast a horizontal 96-sample gel for this purpose. A gel of such large size is difficult to cast, handle, and image, and requires a long run time. These issues can be resolved by using two of Bio-Rads 2 x 32-well ReadyAgarose precast gels instead. ReadyAgarose gels are cast in their own UV-transparent running tray with fluorescent well numbers and a ruler for easy reference. Their gel trays have autolock tabs that snap into the dedicated wide mini ReadySub-Cell GT cell, so you can simply drop one of these precast gels into place and load your samples. ReadyAgarose gels fit most non-Bio-Rad wide mini subcells as well and thus do not require equipment from a specific manufacturer. ReadyAgarose gels are available in 0.8%, 1%, and 3% agarose in TBE or TAE buffer, with or without ethidium bromide, to fit your research requirements. In addition, two 2 x 32-well ReadyAgarose gels offer more wells than a 96-sample handcast gel, run in 3045 min, and have 4.0 cm of resolving distance. These precast gels are designed to give the same results as traditional handcast gels in a fraction of the time.

Hand Casting a 96-Sample Gel
A gel was made by adding 8.7 g of Bio-Rad Certified low range ultra agarose (the same agarose used to manufacture 3% ReadyAgarose gels) to 290 ml of 1x TAE buffer and heating the mixture in a microwave oven to dissolve the agarose. The time required to melt the agarose was noted. The liquid was allowed to cool for 10 min, and 14.5 l of 10 mg/ml ethidium bromide was added. The molten agarose was then poured into a 25 x 15 cm tray enclosed in a gel caster. Two combs, each with 51 wells, were placed in the tray. The agarose was then allowed to solidify and the time for this process was noted.

Preparing and Loading the Samples
Samples were prepared for electrophoresis by mixing 1.5 l of a 500 bp PCR sample, 3.0 l of 5x sample loading buffer containing Orange G, and 10.5 l of 1x TAE buffer for each sample. The samples were loaded into the 102-well handcast gel and two 2 x 32-well, TAE 3% ReadyAgarose gels (with ethidium bromide) using a multichannel pipet. Several lanes on each gel were loaded with Bio-Rads EZ Load 100 bp or 500 bp ruler as reference standards.

Running the Gels
The 102-well gel was run in a Sub-Cell Model 96 cell at 100 V using the PowerPac 300 power supply. The two ReadyAgarose gels were run simultaneously in two wide Mini-Sub Cell GT systems, also at 100 V. TAE buffer (1x) was used to run all of the gels, and the volume of buffer used for each run was noted. In both cases the gels were run until the Orange G dye migrated to 3.5 cm from the well. The running times were noted for both the handcast and precast gels.

Imaging the Gels
Bio-Rads Fluor-S imager and Quantity One 1-D image analysis software were used to image the gels. The handcast gel was imaged as one large gel. The ReadyAgarose gels were imaged as a composite.

Comparative Differences
To run the handcast gel, 2 L of 1x TAE was required, while only 1.2 L of the same buffer was needed to run the two ReadyAgarose gels

It took 45 min to run the ReadyAgarose gels at 100 V, while it took 90 min to run the handcast gel at the same voltage

When the gels were imaged, the background of the handcast gel was not uniform, suggesting that the ethidium bromide was not distributed uniformly, probably because of the high viscosity of a 3% agarose solution (Figure 1). However, no such unevenness was seen in the ReadyAgarose gels (Figure 2)

With the numbered wells on the ReadyAgarose gel tray, documentation of lane samples was both easier and less prone to error. The ruler on the ReadyAgarose tray also facilitated easy and accurate measurement of band migration distances

Time and Cost Differences
The total time needed to prepare and run the handcast and precast gels is compared in Figure 3. A sample cost analysis for 3% gels is presented in Table 1.

In our tests, both handcast and precast gels gave equivalent band sharpness for PCR samples. However, the use of two 2 x 32-well ReadyAgarose precast gels instead of a large 96-sample (102-well) handcast gel saves preparation and running time. Two ReadyAgarose 2 x 32-well gels offer more lanes than a handcast gel designed specifically for 96 samples, thus allowing extra samples or standards to be included in a single run. Data from ReadyAgarose gels are easier to document since well numbers and a ruler are printed on the tray. These advantages of using two ReadyAgarose gels over one large handcast gel make them suitable and desirable for high-throughput laboratories where gel-to-gel consistency is important and relatively small differences in time can accumulate over the long run. Using ReadyAgarose precast gels may also be cost-effective, as this particular comparison demonstrates.

back to top


Page: All 1 2 3 4

Related biology technology :

1. Advantages of Roche Applied Science amplification products
2. SeeSNP Propietary Analysis Advantages
3. CastAway Precast Gels for Rapid Automated DNA Sequence Analysis
4. Four New CastAway Precast Gels
5. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
6. Highest Possible Transformation Efficiencies for High-Throughput Applications
7. High-Throughput Cultivation of Bacterial Clones
8. High-Throughput Isolation of Total RNA
9. High-Throughput System Generates Ultra-Pure PCR Products Suitable for All Applications
10. The DIG System Nonradioactive Automated High-Throughput In Situ Hybridization: a Powerful Tool for Functional Genomics Research
11. Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A
Post Your Comments:

(Date:11/24/2015)... ... November 24, 2015 , ... The United States Golf Association (USGA) ... USGA Green Section Award. Presented annually since 1961, the USGA Green Section Award recognizes ... with turfgrass. , Clarke, of Iselin, N.J., is an extension specialist of ...
(Date:11/24/2015)... Florida (PRWEB) , ... November 24, 2015 , ... ... biggest event of the year and one of the premier annual events for ... and ran from 8–11 November 2015, where ISPE hosted the largest number of ...
(Date:11/24/2015)... ... November 24, 2015 , ... The Academy of Model Aeronautics ... (SIG), MultiGP, also known as Multirotor Grand Prix, to represent the First–Person View (FPV) ... Many AMA members have embraced this type of racing and several new model aviation ...
(Date:11/24/2015)... , Nov. 24, 2015  Twist Bioscience, ... that Emily Leproust, Ph.D., Twist Bioscience chief executive ... Healthcare Conference on December 1, 2015 at 3:10 ... in New York City. --> ... . Twist Bioscience is on Twitter. Sign ...
Breaking Biology Technology:
(Date:11/16/2015)... , Nov 16, 2015  Synaptics Inc. ... human interface solutions, today announced expansion of its ... ™ touch controller and display driver integration ... of smartphones. These new TDDI products add to ... (HD resolution), TD4302 (WQHD resolution), and TD4322 (FHD ...
(Date:11/12/2015)... Mass. , Nov. 12, 2015  Arxspan ... Institute of MIT and Harvard for use of ... discovery information management tools. The partnership will support ... both biological and chemical research information internally and ... will be used for managing the Institute,s electronic ...
(Date:11/11/2015)... 11, 2015   MedNet Solutions , an innovative SaaS-based ... research, is pleased to announce that it will be a ... event, to be held November 17-19 in ... live demonstrations of iMedNet , MedNet,s easy-to-use, ... iMedNet has been able to deliver time and cost ...
Breaking Biology News(10 mins):