Navigation Links
Total RNA Purification from Cultured Cells Using the ABI PRISM 6700 Automated Nucleic Acid Workstation and Total RNA Lysis Reagents

workstation.

5.Activate the default RNA archive protocol to purify total RNA from the cell lysate (Figure 3).

6.If the default RNA Archive protocol is used, purified RNA will be eluted in 150 μL of PCR compatible buffer. Note: If the cell culture growth media is Ca2+/Mg2+ free, lysis may be performed directly in the growth media by addition of an equal volume of 2X lysis reagent.

Results

Applied Biosystems total RNA lysis chemistry is specified to give the following results when used as recommended.

Yield of Total RNA (RNase-free) Total RNA recovery: > 50% for seeding densities in the range of 104 and106 cells per well (Figure 4 and 5)

This recovery rate may be routinely assessed using the Cell Lysate Control Kit, containing lysates at two concentrations: 104 and 106 cells/well. Total RNA is purified from these two lysates and compared to RNA standards included in the kit in an ABI PRISM 7700 Sequence Detection System based RT+ 18S ribosomal RNA assay.

Precision and Reproducibility of Recovery Coefficient of variation (%CV): <30%

Total experimental coefficient of variation, including 6700 workstation purification, all pipetting steps and analysis by ABI PRISM 7700 Sequence Detection System is less than 30% (Figure 4 and 6).

Purity of Total RNA

Purity: < 0.5% w/w of genomic DNA

Demonstrated performance of A260/280 ratio: >1.9 (Figure 6 and 7)

Inhibition

CT suppression of neat sample: ≤0.5

The presence of inhibitors after purification of total RNA samples is a widespread problem with other total RNA extraction methods. The ABI PRISM 7700 Sequence Detection System and an RT+ GAPDH based assay easily
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. New Kit Generates Exceptionally Pure Total RNA for RT-PCR
2. High-Throughput Isolation of Total RNA
3. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
4. Fast Isolation of Total RNA from Small Numbers of Cells
5. Total RNA Isolation Protocol 1,2,3
6. Isolate It All: siRNA miRNA Total RNA Native Protein
7. Isolate Total RNA and Protein From the Same Sample
8. Isolate High Quality Total RNA from LCM Samples Suitable for Microarray and qRT-PCR Analysis
9. Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
10. Total RNA from Whole Blood for Expression Profiling
11. RNAPrep - Total RNA Isolation Purification
Post Your Comments:
(Date:1/22/2015)... | RBJ today announces the firm brokered a long-term lease ... leading biopharmaceutical company, at Two Ledgemont Center in ... , president, and Brian Cohen , senior vice president, ... at 95 Hayden Ave. Photo - http://photos.prnewswire.com/prnh/20150122/170765 ...
(Date:1/22/2015)... BOSTON , Jan. 22, 2015  Transwestern | RBJ today ... feet of office space for Shire a leading biopharmaceutical company, ... . Transwestern | RBJ,s Robert Richards , president, and ... the lease for the entire five-floor building at 95 Hayden ...
(Date:1/22/2015)... 2015 The laboratory information management systems ... a number of technological advancements due to factors such ... need to integrate healthcare systems, and increasing government support ... system integration. Key players in the market focus on ...
(Date:1/22/2015)... Rootstown, OH (PRWEB) January 22, 2015 Crystal ... announced today that it has received AOAC-PTM Certifications for the ... O111, O121, and O145; collectively referred to as STEC or ... at 1 colony forming unit (cfu) per 325 g of ...
Breaking Biology Technology:Transwestern | RBJ Advises Shire in 202,000 SF Lease, Creating Boston's Largest Suburban Biotech Campus 2Transwestern?RBJ Advises Shire in 202,000 SF Lease, Creating Boston's Largest Suburban Biotech Campus 2Laboratory Information Management Systems Market is Expected to Reach $1,323.6 Million by 2019 - New Research Report by MarketsandMarkets 2Laboratory Information Management Systems Market is Expected to Reach $1,323.6 Million by 2019 - New Research Report by MarketsandMarkets 3Laboratory Information Management Systems Market is Expected to Reach $1,323.6 Million by 2019 - New Research Report by MarketsandMarkets 4Crystal Diagnostics Awarded AOAC-PTM Accreditation for the Rapid Detection of “Big 6” E.coli Food Pathogens 2
... Cord Blood America, Inc . ( www.cordblood-america.com ) ... blood stem cell preservation company focused on bringing ... and internationally, today announced that revenues for 2011 increased 37 ... The Company announced a gross profit increase in 2011 of ...
... 2012   HemoShear, LLC , a biotechnology research company ... systems for discovery and assessment of new drug compounds, ... Heart Association,s Arteriosclerosis, Thrombosis and Vascular Biology (ATVB) conference ... at the Hilton Chicago, 720 South Michigan Avenue in ...
... A recent solicitation by the Defense Logistics Agency ... a deluge of orders for emergency radiation products supplier ... tablets provide protection against radioactive iodine in the event ... of nuclear weaponry in Asia and the Middle East ...
Cached Biology Technology:Cord Blood America Revenues Up 37 Percent to $5.67 Million in 2011 2HemoShear to Present at Arteriosclerosis, Thrombosis and Vascular Biology (ATVB) Conference 2Potassium Iodide Sales Surge on Heels of Large Government Order 2
(Date:12/22/2014)... , Dec. 22, 2014  NXT-ID, Inc. (NASDAQ: NXTD ... company focused on the growing mobile commerce market, announces it ... Robbins for the 2015 International CES debut of the ... will be at the NXT-ID booth January 6th and ...
(Date:12/19/2014)... Research and Markets ( http://www.researchandmarkets.com/research/8kmfcn/iphone_5s ) has ... Sensor - Apple/AuthenTec TMDR92 & Sapphire - Technology Report" ... http://photos.prnewswire.com/prnh/20130307/600769 Following the acquisition of ... feature with the iPhone 5S. It is currently the ...
(Date:12/19/2014)... Dec. 18, 2014   LaunchKey , the first decentralized ... and Internet of Things era, today announced the close ... round was led by Metamorphic Ventures with participation from ... VegasTechFund, and others.  LaunchKey has raised $4 million to ...
Breaking Biology News(10 mins):NXT-ID Hires Master Pickpocket and Security Consultant Apollo Robbins for Wocket Smart Wallet CES 2015 Debut 2NXT-ID Hires Master Pickpocket and Security Consultant Apollo Robbins for Wocket Smart Wallet CES 2015 Debut 3NXT-ID Hires Master Pickpocket and Security Consultant Apollo Robbins for Wocket Smart Wallet CES 2015 Debut 4iPhone 5S Fingerprint Sensor - Apple/AuthenTec TMDR92 & Sapphire - Technology Report 2LaunchKey Raises $3 Million in Additional Funding Led by Metamorphic Ventures 2LaunchKey Raises $3 Million in Additional Funding Led by Metamorphic Ventures 3
... diseases such as schizophrenia, major depression and cancer ... of dysfunctional genes. A novel computational biology method ... Abdi, PhD, http://www.njit.edu/news/2008/2008-367.php , associate professor in ... found a way to uncover the critical genes ...
... day Americans elect a new president, one of the nation,s ... Houston and outline the daunting global warming challenges awaiting the ... analysis at the National Center for Atmospheric Research (NCAR) in ... a colloquium Tuesday, Nov. 4. The event is free ...
... granted to the laboratory of William Roush, Ph.D., ... Research Institute. Other participants in the consortium are ... California, San Francisco (UCSF), and University of Washington. ... Translational Research Institute Medical Chemistry division and professor ...
Cached Biology News:NJIT professor finds engineering technique to identify disease-causing genes 2Climate change expert to discuss grim global warming predictions 2Scripps Florida scientists awarded $1.5M to fight major water and food parasites 2
... can provide custom microplate coating using advanced automated high throughput ... plate formats. We can also accommodate virtually any lot size ... L):, , 8 and ... microplate, 384 well microplate, ...
...
A flexible 24-48 hour DNA sequencing service. Samples may be shipped in standard 1.5mL tubes. Oligo synthesis and DNA preparation available. Read lengths up to 850 Phred 20 bases. Universal primers p...
... automated, GLP-compliant Facility , Galaxy LIMS, Sophisticated ... Highly experienced scientists ,We offer individual sample ... full plasmids for single or double strand ... when the DNA of interest is 1.3 ...
Biology Products: