Navigation Links
Your Data: Detection of Oocyte-specific Gene Expression Using Ambion's Cells-to-cDNA Kit


Ambion's Cells-to-cDNA Kit produces cDNA ideal for RT-PCR based gene quantitation, including experiments performed using real-time RT-PCR. The Cells-to-cDNA Kit comes in an easy-to-use format that produces first-strand cDNA from cultured cells without RNA isolation in less than 2 hours. By using an RT-compatible Cell Lysis Buffer that inactivates RNases, RT-PCR can be carried out on only a few cells.

Using the Cells-to-cDNA Kit, Dr. Alex Bortvin from the Whitehead Institute (Cambridge, Massachusetts) has successfully carried out RT-PCR amplification of an oocyte-specific gene and a housekeeping gene from individual mouse oocytes. Dr. Bortvin made some modifications to the Cells-to-cDNA protocol for this application. One fertilized oocyte (in 1 l of culture medium) was added to 5 l of Cell Lysis Buffer and overlaid with oil to prevent evaporation. After cell lysis, DNase I treatment was carried out with a 1:10 dilution of the enzyme at 37C for 30 min. cDNA was synthesized according to the Cells-to-cDNA protocol.

In order to ensure that clean templates for cDNA library synthesis and multiplex PCR applications was obtained, Dr. Bortvin purified the first-strand cDNA using a PCR purification kit (note that this should not normally be necessary). This step enabled him to use the PCR components and conditions that he had previously optimized for his gene of interest. PCR was carried out with gene-specific primers for 50 cycles. A "minus-RT" control reaction was included to rule out the presence of any trace genomic DNA contamination; none was seen (data not shown).


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Your Data: Use of pSilencer 1.0-U6 siRNA Expression Vector in the Analysis of Synaptotagmin Location and Function
2. Your Data: RNAlater Stories
3. Your Data: Silencing Hepatitis C Virus Replication With In Vitro Transcribed siRNAs
4. Your Data: Silencing Neural Cell Adhesion Genes with In Vitro Transcribed siRNAs
5. Your Data: Silencing of a Pro-apoptotic Transcription Factor Using Synthetic siRNA
6. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
7. A New PCR-based Mycoplasma Detection Method
8. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
9. Detection and Identification of Phosphorylation Sites in Proteins Using LC/MS/MS with Neutral Fragment Loss Mapping
10. Detection of mRNAs on Cryosections of the Cardiovascular System Using DIG-Labeled RNA Probes
11. Gene Expression Arrays: Highly Sensitive Detection of Expression Patterns with Improved Tools for Target Amplification
Post Your Comments:
(Date:8/29/2014)... Local veterinarian, Dr. Keith Clement, and ... candidates to participate in an investigational study of donor ... goal of this study is to determine if a ... two arthritically affected joints can help reduce pain and ... the current investigational study must be older than nine ...
(Date:8/29/2014)... 29, 2014 Due to a misstatement posted ... the Adult Stem Cell Technology Center, LLC ( ASCTC ) ... cell therapy research that might benefit ALS patients. Because ... new stem cell technologies and therapeutic applications, FRC listed ASCTC ... Challenge donors who wish not to support research that ...
(Date:8/29/2014)... Antonio, TX (PRWEB) August 29, 2014 ... and ISO 13485:2003 certified, GAMP® 5 compliant imaging ... awarded an international Phase II clinical trial to ... lymphoma. , Throughout this trial, Intrinsic Imaging will ... not limited to, protocol and charter development, site ...
(Date:8/28/2014)... Best Sanitizers, Inc., a leader in hand ... is asking industry professionals to prepare for fall harvest ... soap they’re currently using to the Best Sanitizers’ ... hygiene is critical to fighting cross-contamination and the spread ... criteria are identified to evaluate the effectiveness of an ...
Breaking Biology Technology:Burnt Hills Veterinary Hospital Seeks Candidates for an Investigational Study of Stem Cells for Dogs with Arthritis 2The Adult Stem Cell Technology Center, LLC Declines ALS Ice Bucket Challenge Donors Wishing to Support Ethical Research for New Stem Cell Therapies 2The Adult Stem Cell Technology Center, LLC Declines ALS Ice Bucket Challenge Donors Wishing to Support Ethical Research for New Stem Cell Therapies 3Intrinsic Imaging Awarded Phase II Clinical Trial to Assess New Treatment for Non-Hodgkin’s Lymphoma 2Best Sanitizers, Inc. Asks Food Industry Professionals: With Fall Harvest On the Way, Is Your E2 Hand Soap Up to the Task? 2
... made tremendous scientific advances by dissecting apart the ... at the University of Wisconsin-Madison College of Engineering ... the pieces back together, mathematically. , John Yin, ... computer models of a relatively simple virus to ...
... SCOTTSDALE, Ariz., Feb. 6 Cancer Clinics of ... community oncologists, announced the completion of its 62nd ... leading oncology disease states at the 4th Annual ... oncology group is now fully prepared to launch ...
... LLC, a clinical-stage drug development company commercializing first-in-class ... completion of a $2.9 million private placement. ... and clinical programs for its lead drug compound, ... David G. Kalergis, Chief Executive Officer of Diffusion ...
Cached Biology Technology:Mathematical models reveal how organisms transcend the sum of their genes 2Cancer Clinics of Excellence (CCE) Releases 62nd Evidence-Based Treatment Protocol 2Diffusion Pharmaceuticals Announces Completion of a $2.9 Million Private Placement and Company Presentations at Upcoming Industry Conferences in Geneva, Switzerland and Naples, Florida 2Diffusion Pharmaceuticals Announces Completion of a $2.9 Million Private Placement and Company Presentations at Upcoming Industry Conferences in Geneva, Switzerland and Naples, Florida 3
(Date:9/1/2014)... Boston, MA ─ Dietary quality in the U.S. has ... reduced trans fat intakebut overall dietary quality remains poor ... groups, according to a new study from Harvard School ... most direct evidence to date that the extensive efforts ... quality are having some payoff, but it also indicates ...
(Date:9/1/2014)... Conventional wisdom has long held that corals ... reefs are passive organisms that rely entirely ... as nutrients and oxygen. But now scientists at ... in Israel have found that they are far ... into turbulent patterns that greatly enhance their ability ...
(Date:9/1/2014)... It may be possible to train the brain ... according to new research by scientists at the Jean ... HNRCA) at Tufts University and at Massachusetts General Hospital. ... Diabetes , a brain scan study in adult men ... the addictive power of unhealthy food while also increasing ...
Breaking Biology News(10 mins):Quality of US diet shows modest improvement, but overall remains poor 2Quality of US diet shows modest improvement, but overall remains poor 3Nature's tiny engineers 2Nature's tiny engineers 3Training your brain to prefer healthy foods 2
... gone underground to get the most top secret information ... in his experimental orchards, and it is sort of ... is part of a national specialty crop research project ... Texas AgriLife Research horticultural scientist, will focus on pecans ...
... causes temperatures to increase in Hawaii,s mountains, deadly ... the mountains, invading most of the last disease-free ... and remarkable birds. A just-published U.S. ... a forthcoming "disease invasion" by examining the present ...
... around the world will convene at the University of ... from the most ambitious effort ever undertaken to measure ... they contribute in the near term to the dramatic ... two-year international field campaign known as POLARCAT was conducted ...
Cached Biology News:Getting to the root of science in a nutty way 2Jeepers creepers: Climate change threatens endangered honeycreepers 2Jeepers creepers: Climate change threatens endangered honeycreepers 3Spring agricultural fires have large impact on melting Arctic 2Spring agricultural fires have large impact on melting Arctic 3
p27 rabbit polyclonal antibody...
... Rabbit polyclonal to Ki67 - Proliferation Marker, ... cell cycle related nuclear protein, expressed by ... active cell cycle (G1, S, G2 and ... (G0) cells. Ki67 antibodies are useful in ...
UBC13 Antibody...
Request Info...
Biology Products: