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

fied 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.

Lysis and Purification of RNA from Adherent Cell Cultures

Important: Lysis reagent is formulated as a 2X solution. The final lysis buffer concentration in any lysis step should always be 1X for maximum RNA recovery.

Method 1

Automated lysis reagent addition by the 6700 workstation

1.Remove the growth media by aspiration from all wells.

2.Re-suspend the cells in 125 μL PBS (phosphate buffered saline, Ca2+/Mg2+ free).

3.Place the cell culture plate containing the cells in the primary input position of the 6700 workstation.

4.Activate the default lysis protocol (Figure 2).

5.Configure the lysis protocol to add 125 μL of 2X lysis reagent and to mix the sample at least 3 times after addition.

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

7.If the default RNA archive protocol is used, purified RNA will be eluted in 150 μL of PCR compatible buffer.

Method 2

Manual lysis of adherent cells

1.Remove the growth media by aspiration from all wells.

2.Re-suspend the cells in 250 μL of a 1:1 mixture of lysis reagent and PBS (phosphate buffered saline, Ca2+/Mg2+ free).

3.Ensure complete nucleic acid release by taking up the lysate into the pipette tip at least three times and thoroughly mixing the lysis buffer with the cells.

4.Place the cell culture plate containing the lysate in the primary input position of the 6700


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:5/28/2015)... MARLBOROUGH, Mass. , May 28, 2015 ... company focused on discovering and developing innovative therapeutics, primarily ... the pricing of a public offering of 26 million ... per share for gross proceeds of approximately $10.4 million. ... stock, (ii) a 13-month overallotment purchase right to purchase ...
(Date:5/28/2015)... GAITHERSBURG, Md. , May 28, 2015  GenVec, ... president and CEO, Douglas J. Swirsky , will ... Healthcare Conference at 11 a.m. EDT on Thursday, June ... webcast of Mr. Swirsky,s presentation will be available at ... also be available on GenVec,s website.  To access, visit ...
(Date:5/28/2015)... 2015 The Fertility Centers of New ... Danielle Vitiello has taken on a new and expanded ... will provide guidance, leadership, oversight and quality assurance for ... Vitiello among our team of experienced reproductive endocrinologists,” said ... A. Hill, M.D. “Her expertise and compassionate, individualized care ...
(Date:5/28/2015)... Whitehouse Laboratories is excited to announce that ... with PTI Inspection Systems. Whitehouse Labs and PTI will ... method development and validation programs using the most ... includes a more formal program that facilitates cross training ... enables a more effective team approach, especially as it ...
Breaking Biology Technology:RXi Pharmaceuticals Announces $10 Million Public Offering of Common Stock and Warrants 2RXi Pharmaceuticals Announces $10 Million Public Offering of Common Stock and Warrants 3GenVec To Present At The Jefferies Healthcare Conference 2Danielle Vitiello, MD of Fertility Centers of New England, Named New Medical Director 2Whitehouse Labs Renews & Enhances Partnership with PTI Inspection Systems 2Whitehouse Labs Renews & Enhances Partnership with PTI Inspection Systems 3
... Milwaukee, Wis. - In a move to comply ... PassMark Security, an Internet security provider, as a partner ... credit union clients. , ,The move comes as financial ... to comply with Federal Financial Institutions Examination Council requirements ...
... McGlenn kept busy at the BIO Chicago industry show this ... a laser-pointer pen or kangaroo lapel pin, he met with ... grow mostly in prearranged sessions designed to help attendees ... is CEO of NeoClone , which manufactures monoclonal antibodies. ...
... are good for the UK-based parent company of ... revenues during the first quarter of 2006. , ,Profits ... GE Healthcare Technologies, Waukesha, and GE Healthcare Bio-Sciences, rose ... operating income grew to $496 million from $409 million. ...
Cached Biology Technology:Partner program is highlight of BIO for Wisconsin firm 2
(Date:5/20/2015)... 2015 NXT-ID, Inc. (NASDAQ: NXTD ) ... subsidiary, 3D-ID LLC, a company engaged in biometric identification ... biometrics technology portion of the recently awarded TIES (Technical ... of the Army. 3D-ID LLC has a ... law enforcement and security agencies. Gino ...
(Date:5/18/2015)... , May 18, 2015 Fingerprint Cards ... FPC1020, FPC1025, FPC1145, FPC1155 and FPC1035 from one if its ... until and including Q3 2015 and the sensors will be ... Communicated order values for 2015 hereby amount to 740 MSEK ... 2015 of 140 MSEK and a number of smaller orders ...
(Date:5/11/2015)... 11, 2015  Through a well-rounded UAS delegation representing private ... a strong showing at AUVSI,s Unmanned 2015 conference last week ... Ohio,s UAS industry met with over 200 ... points along the UAS ecosystem. "Our message ... President for Aerospace Rich Knoll . "If you want ...
Breaking Biology News(10 mins):NXT-ID, Inc Announces Participation as Member of Team Battelle for TIES Contract Award 2NXT-ID, Inc Announces Participation as Member of Team Battelle for TIES Contract Award 3Fingerprint Cards Receives Touch Fingerprint Sensor Order of SEK 230 Million and Raises Revenue Guidance for 2015 2Ohio Flies High at Unmanned '15, Sets Stage for Ohio UAS Conference 2
... When it comes to understanding a critical junction in ... real thorn in scientists, sides. Specialists have jousted over ... called Acoelomorpha . This collection of worms, which ... larger group called bilateral animals, organisms that have symmetrical ...
... The European College of Neuropsychopharmacology (ECNP) ... recipient of the 2009 ECNP Neuropsychopharmacology Award in ... and innovative research into the neurochemical basis of ... recognises outstanding research in the field of neuropsychopharmacology ...
... IEEE-USA President Gordon Day said today that he strongly ... White House and discussed by President Barack Obama in ... Troy, N.Y. "IEEE-USA recognizes the vital role that ... U.S. competitiveness, energy independence and national security," Day said. ...
Cached Biology News:Research team finds first evolutionary branching for bilateral animals 2
... Conjugate, 2 ml. Streptavidin ... molecules for use in ... hybridization applications.High specificity reagents. ... & Detection, Protein Labeling ...
... membrane-filtered isotonic and enzyme-free aqueous formulations ... agents in either Ca2+- and Mg2+-free ... Ca2+- and Mg2+-free phosphate-buffered saline for ... support substrates and each other for ...
... Cell Dissociation Buffers are membrane-filtered ... salts chelating agents and cell-conditioning agents ... balanced salt solution or Ca2+- ... dissociation of mammalian cells from support ...
Biology Products: