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:8/27/2015)... , Aug. 27, 2015 /PRNewswire/ - Portage Biotech ... PBT.U), and Biohaven Pharmaceutical Holding Company Limited (Biohaven), ... Phase I study of BHV-0223, a glutamate modulating ... regarding BHV-0223 and recently obtained clearance from the ... with human testing. Portage holds 54% equity interest ...
(Date:8/27/2015)... , August 27, 2015 GlassesOff ... image processing functions in the human vision system, announced today ... point guard Chris Paul to develop a ... who want to improve their real-life on-court performance. ... court, whether it is shooting a ball or blocking a ...
(Date:8/26/2015)... ... August 26, 2015 , ... ... today that a second US laboratory is to open in Manhattan, Kansas in ... continued partnership and long-term growth of research and development through collaboration with researchers ...
(Date:8/26/2015)...  Platform therapeutic company Symic Biomedical, Inc. ("Symic") ... Phase II SBIR grant from the National Institutes ... agent to reduce arteriovenous fistula (AVF) failures, a ... disease (ESRD) patients undergoing hemodialysis. The two-year project, ... Digestive and Kidney Diseases (NIDDK), will allow Symic ...
Breaking Biology Technology:Portage's Biohaven doses first human subject in Phase 1 trial with lead drug candidate BHV-0223 2Portage's Biohaven doses first human subject in Phase 1 trial with lead drug candidate BHV-0223 3Portage's Biohaven doses first human subject in Phase 1 trial with lead drug candidate BHV-0223 4Eight-Time All-Star Chris Paul and GlassesOff Partner to Develop Innovative Mobile App Designed to Improve On-Court Performance 2Eight-Time All-Star Chris Paul and GlassesOff Partner to Develop Innovative Mobile App Designed to Improve On-Court Performance 3MediVet Biologics Animal Health Secures Additional Laboratory Facility in Manhattan, Kansas 2MediVet Biologics Animal Health Secures Additional Laboratory Facility in Manhattan, Kansas 3Symic Biomedical Awarded $1.5MM NIH Phase II SBIR Grant to Develop its Proteoglycan Mimetic Therapeutics to Reduce Vascular Access Failures in ESRD 2Symic Biomedical Awarded $1.5MM NIH Phase II SBIR Grant to Develop its Proteoglycan Mimetic Therapeutics to Reduce Vascular Access Failures in ESRD 3
... Recurring Disposables, Services and Accessories Revenue ... Irrigated Catheter Launch Provided, ST. LOUIS, March 3 ... the fourth quarter and full year,ended December 31, 2007., ... to $39.3 million,compared with $27.2 million in 2006. Sales ...
... Feb. 29 For the first time,researchers at ... spheres whose halves are physically or chemically,different -- ... when an alternating electrical field is applied to ... research, published in the Feb. 8, 2008, edition,of ...
... Feb. 29 Kendle (Nasdaq: KNDL ... today,announced that Vice President and Chief Marketing ... Annual Raymond James,Institutional Investors Conference. The conference ... Cypress in Orlando, March 2-5, 2008. Kendle,s,presentation ...
Cached Biology Technology:Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 2Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 3Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 4Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 5Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 6Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 7Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 8Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 9Stereotaxis Reports Fourth Quarter and Full Year 2007 Financial Results 10'Two-Faced' Particles Act Like Tiny Submarines 2'Two-Faced' Particles Act Like Tiny Submarines 3Kendle to Present at the 29th Annual Raymond James Institutional Investors Conference 2
(Date:8/5/2015)... August 5, 2015 According to ... Management Market by Software, Services, Vertical (BFSI, Airport, IT & ... - Global Forecast to 2020", published by MarketsandMarkets, defines ... globally into various segments. The global PIAM Market is ... $546.2 Million by 2019, at a CAGR of 14.9 ...
(Date:8/4/2015)... Aug. 04, 2015 ... has announced the addition of the ... and Service), Sub-Segment (Computer Forensics, Network Forensics, ... Types, Service, Vertical and Region - Global ... offering. By Component (Hardware, Software, ...
(Date:7/31/2015)... Kina, 31. juli 2015 Den 10. internationale ... af BGI fra den 22. - 25. oktober i ... fejrer sin 10-års fødselsdag i år. Siden starten i ... årlige møder på ,omik,-feltet, og er en af de ... ICG-10 fokuserer på nylige gennembrud og fremskridt inden ...
Breaking Biology News(10 mins):Physical Identity and Access Management Market Worth $546.2 Million by 2019 2Physical Identity and Access Management Market Worth $546.2 Million by 2019 3Physical Identity and Access Management Market Worth $546.2 Million by 2019 4Digital Forensics Market - The Market Represents a Compound Annual Growth Rate (CAGR) of 11.3% From 2015 to 2020 2Den 10. internationale konference om Genomics (ICG-10) åbner i oktober 2
... known that racing cars and bicyclists can reduce air ... from New York University and Cornell University have found ... flags. Their study, published in the most recent issue ... that in a series of flags, the leading flag ...
... increasing popularity of reduced tillage on crops has not ... but it has also been associated with increasing organic ... peanut crops, however, reduced tillage has not gained a ... inconsistent yields have not encouraged farmers to make a ...
... With childhood obesity increasing, school administrators and public health ... schools. In a study published in the November/December 2008 ... Behavior , researchers found that reduction or elimination of ... consumption by adolescents. Working with four schools ...
Cached Biology News:Lead-flapping objects experience less wind resistance than their trailing counterparts 2Tillage, rotation impacts peanut crops 2
... LiCl precipitation offers major advantages over other ... precipitate DNA, protein or carbohydrates. It is the ... or cDNA synthesis from RNA preparations. Note, LiCl ... total RNA isolations. If the RNA is to ...
...
...
The Captivate Disposable Sample Chamber is designed for use with the Captivate magnetic yoke assembly (C-24700) and can be used with our Captivate ferrofluid conjugates (C-21473 C-21474 C-21476) or ...
Biology Products: