Navigation Links
Automating RNA Isolation

lity on an Agilent 2100 bioanalyzer. Yields were consistently high (~5 pg per cell for HeLa S3 cells and ~8 pg per cell for K562 cells) and RNA samples were uniformly intact. Furthermore, genomic DNA was effectively removed. An electropherogram demonstrating the high quality of the RNA recovered is shown in Figure 1.

Figure 1.RNA Isolated Using the Automated RNAqueous-96 Protocol. Cultured K562 cells (human leukemia, grown in suspension) were pelleted and resuspended in 1X phosphate buffered saline (PBS), counted and transferred to 48 wells of a deep well 96-well plate at 3.75 x 105 cells/well. Total RNA was isolated using the automated method described at "RNAqueous-96 Automation Protocol", using RNAqueous-96 and a MultiPROBE II HT Liquid Handling System with a Gripper Integration Platform. This RNA sample, which was representative of the 48 samples isolated and analyzed, has a 28S rRNA to 18S rRNA ratio of 1.92, indicating a high level of integrity. This isolation yielded 24 g of RNA (344 ng/l in 70l).


Lack of reproducibility can be an issue with some RNA isolation methods. Real-time RT-PCR was used to assess the reproducibility of the automated RNAqueous-96 procedure. Because real-time RT-PCR is quantitative, reproducible, and can be used to analyze multiple samples simultaneously, it is a good choice for such an analysis. Both an abundant (GAPDH) and a rare target (hTERT: catalytic subunit of human telomerase) were individually analyzed in the previously described HeLa S3 and K562 RNA samples using a TaqMan assay. R
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
2. High-Throughput Isolation of Total RNA
3. Simple Isolation of RNA from Tissue and Cultured Cells
4. Fast Isolation of Total RNA from Small Numbers of Cells
5. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
6. Isolation of Microorganisms
7. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
8. Mouse Tail Genomic DNA Isolation Protocol(1)
9. Genomic DNA Isolation Protocol(1,2,3)
10. Basic Plasmid DNA Isolation Protocol
11. Protocol for RNA Isolation Using TRIzol Reagent with Phase Lock Gel-Heavy
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:5/24/2017)... ... May 24, 2017 , ... Patient ... developed with Wi-Fi connectivity to reduce the amount of wiring in a healthcare ... addition, compact mobile devices including infusion pumps, heart and hypertension monitoring, glucose monitoring, ...
(Date:5/23/2017)... ... 2017 , ... Firmex today announced the general ... for organizations to send and gather large files and confidential documents beyond the ... size limitations. , Using the same market-tested infrastructure as Firmex’s flagship Virtual ...
(Date:5/23/2017)... ... May 23, 2017 , ... ... source of human cardiovascular cells for research and the development of cardiac ... possible to generate large numbers of cardiomyocytes (hPSC-CMs). Due to varying differentiation ...
(Date:5/23/2017)... ... May 22, 2017 , ... NetDimensions has been ranked as ... Globe™ for Corporate Learning, 2017. , Aragon Research defines Leaders as organizations who ... perform against those strategies. NetDimensions’ ranking as a Leader due to its strengths ...
Breaking Biology Technology:
(Date:4/5/2017)... Today HYPR Corp. , leading innovator ... of the HYPR platform is officially FIDO® Certified ... architecture that empowers biometric authentication across Fortune 500 enterprises ... over 15 million users across the financial services industry, ... product suites and physical access represent a growing portion ...
(Date:4/3/2017)...  Data captured by IsoCode, IsoPlexis Corporation,s ... statistically significant association between the potency of ... objective response of cancer patients post-treatment. The ... cancer patients will respond to CAR-T cell ... to improve both pre-infusion potency testing and cell ...
(Date:3/30/2017)... Trends, opportunities and forecast in this market ... (fingerprint, AFIS, iris recognition, facial recognition, hand geometry, vein ... use industry (government and law enforcement, commercial and retail, ... others), and by region ( North America ... Pacific , and the Rest of the World) ...
Breaking Biology News(10 mins):