Navigation Links
Electroporation of Yeast Artificial Chromosomes

Maren Bell and Robert Mortimer, Human Genome Center, Division of Cell and Molecular Biology, Lawrence Berkeley Laboratory, Berkeley, California 94720


Introduction
Transformation of yeast with yeast artificial chromosomes (YACs) has traditionally been performed by a PEG-spheroplast procedure.1,2 However, the procedure is complicated, often unreliable, and transformation efficiencies are relatively low when compared to more recently developed transformation methods. An additional concern is that the smaller YACs of any given ligation mixture are cloned preferentially, therefore prior size fractionation with sucrose gradients to enrich for larger molecules, or treatment with spermidine to improve transformation efficiency of larger molecules, is required to obtain reasonable efficiencies with larger YACs. Several electroporation protocols have recently been reported for the transformation of Saccharomyces cerevisiae with supercoiled plasmids.3-6 In an attempt to improve the currently used YAC cloning procedure, we investigated the electroporation of S. cerevisiae strain AB1380 (Mat a, ade2-1, can1-100, lys2-1, trp1, ura3, his5, psi + ) with a 14.4 kb mini-YAC. Initial experiments showed transformation efficiencies two-fold better than those reported in a previously published YAC electroporation protocol.7


Materials and Methods
The YAC used in our experiments was constructed by the insertion of a 4.6 kb lambda MluI fragment into the MluI-SUP4 cloning site of pYAC-RC. 8 E. coli was transformed by the resulting 16.1 kb plasmid, the DNA was isolated, purified over cesium chloride, and restriction d
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Eppendorf Electroporation Application Notes Your participation is highly welcome !!!
2. Eppendorf Electroporation Application Notes Your participation is highly welcome !!!
3. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
4. Optimizing Chemical Transfection and Electroporation of siRNAs
5. High Throughput siRNA Electroporation
6. Electroporation of Primary Bone Marrow Cells
7. Production of Hybridomas by Electroporation
8. Transformation of Filamentous Fungi by High-Voltage Electroporation
9. Transgenic Zebrafish by Electroporation
10. Transient Gene Expression Following Single and Double Exponential Decay Electroporation Pulses
11. Epitope-Tagging Vectors for Functional Analysis in Yeast
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Electroporation Yeast Artificial Chromosomes

(Date:7/3/2015)... ... July 03, 2015 , ... The Alexander Group, a ... service industries alike-- typically known for its consummate professionalism-- embarks each year in ... and break from their day jobs. , "Our motto has always been work ...
(Date:7/3/2015)... -- Pomerantz LLP announces that a class action lawsuit has ... "Company")(NYSE: PBYI ) and certain of its officers.   ... Central District of California , and ... consisting of all persons or entities who purchased Puma ... inclusive (the "Class Period").  This class action seeks to ...
(Date:7/2/2015)... Md. , July 2, 2015  Spherix Incorporated ... to the fostering and monetization of intellectual property, today ... of Spherix Incorporated v. Verizon Services Corp. et ... District Court for the Eastern District of ... on July 1, 2015 interpreting certain key claims in ...
(Date:7/1/2015)... ... July 01, 2015 , ... It is now possible to separate two common ... emission computed tomography (SPECT) according to researchers from Amen Clinics, UCLA, Thomas Jefferson ... scans and is believed to be the largest brain imaging study ever, will be ...
Breaking Biology Technology:The Alexander Group Declares Their Independence with a Bit of Levity 2SHAREHOLDER ALERT: Pomerantz Law Firm Reminds Shareholders with Losses on their Investment in Puma Biotechnology, Inc. of Class Action Lawsuit and Upcoming Deadline - PBYI 2SHAREHOLDER ALERT: Pomerantz Law Firm Reminds Shareholders with Losses on their Investment in Puma Biotechnology, Inc. of Class Action Lawsuit and Upcoming Deadline - PBYI 3SHAREHOLDER ALERT: Pomerantz Law Firm Reminds Shareholders with Losses on their Investment in Puma Biotechnology, Inc. of Class Action Lawsuit and Upcoming Deadline - PBYI 4Spherix Issues A Litigation Update 2Spherix Issues A Litigation Update 3Brain Scans Can Tell Post-Traumatic Stress Disorder and Traumatic Brain Injury Apart According to New Research by Amen Clinics 2Brain Scans Can Tell Post-Traumatic Stress Disorder and Traumatic Brain Injury Apart According to New Research by Amen Clinics 3Brain Scans Can Tell Post-Traumatic Stress Disorder and Traumatic Brain Injury Apart According to New Research by Amen Clinics 4
... , NEW YORK, ... research report is available in its catalogue: , ... , http://www.reportlinker.com/p0165503/Brazil-Bioethanol-Market-Analysis-and-Forecasts-to-2013.html , Brazil Bioethanol Market Analysis ... The report provides detailed analysis and forecasts of the ...
... ... Ltd (ASX:NEU) has agreed an AU$6.7 mil convertible note private placement with a U.S. ... agent and advisor. , ... New York (PRWEB) November 23, 2009 -- The independent New York investment bank ...
... ... grew by 29,000 jobs in October, and the field remains the only one to show ... health systems are coming closer to having to actually cut medical personnel, instead of just ... (Vocus) November 24, ...
Cached Biology Technology:Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 2Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 3Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 4Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 5Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 6Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 7Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 8Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 9Reportlinker Adds Brazil Bioethanol Market Analysis and Forecasts to 2013 10KTA Capital Arranges an AU$6.7 Million Private Placement by Neuren Pharmaceuticals 2The MedZilla Report for October 2009 - Health Care Employment Grows Again in October Even As Clinics, Specialty Centers Close 2The MedZilla Report for October 2009 - Health Care Employment Grows Again in October Even As Clinics, Specialty Centers Close 3The MedZilla Report for October 2009 - Health Care Employment Grows Again in October Even As Clinics, Specialty Centers Close 4
(Date:6/24/2015)... , June 22, 2015 ... announced the addition of the "Huawei Ascend ... report to their offering. Huawei,s ... to the opposite direction than Apple,s and Samsung,s ... fingerprint sensor developed by Fingerprint Cards, a main ...
(Date:6/23/2015)... June 22, 2015 ... addition of the "Body-Worn Temperature Sensors Market ... and Forecast 2014 - 2020" report to ... analysis of the global body-worn temperature sensors market. ... segmented on the basis of types, care setting, ...
(Date:6/18/2015)... June 18, 2015 This report analyzes the ... the following Product Segments, which includes Immunoassay Analyzers and ... Systems, Radio Immunoassay Systems, and Nephelometric Immunoassay Systems. The ... Canada , Japan , ... , Latin America , and Rest ...
Breaking Biology News(10 mins):Huawei Ascend Mate 7 Fingerprint Sensor - Reverse Costing Analysis 2Body-Worn Temperature Sensors Market - Global Industry Analysis 2Global Immunoassay Systems Market 2Global Immunoassay Systems Market 3Global Immunoassay Systems Market 4Global Immunoassay Systems Market 5Global Immunoassay Systems Market 6Global Immunoassay Systems Market 7Global Immunoassay Systems Market 8Global Immunoassay Systems Market 9Global Immunoassay Systems Market 10Global Immunoassay Systems Market 11Global Immunoassay Systems Market 12Global Immunoassay Systems Market 13Global Immunoassay Systems Market 14Global Immunoassay Systems Market 15Global Immunoassay Systems Market 16Global Immunoassay Systems Market 17Global Immunoassay Systems Market 18Global Immunoassay Systems Market 19Global Immunoassay Systems Market 20Global Immunoassay Systems Market 21Global Immunoassay Systems Market 22Global Immunoassay Systems Market 23Global Immunoassay Systems Market 24Global Immunoassay Systems Market 25Global Immunoassay Systems Market 26Global Immunoassay Systems Market 27Global Immunoassay Systems Market 28Global Immunoassay Systems Market 29
... Scientists have discovered the first gene involved in regulating the optimal ... of slumber, an enigmatic phenomenon that is critical to human physical ... 2009 issue of Science , identified a mutated gene that ... hours of sleep a day rather than the eight to eight-and-a-half ...
... Porcine circovirus associated diseases cost pig producers around the ... That,s why Kansas State University virologists Dick Hesse and ... to tackle such complex diseases. "Circovirus and its ... said. "Both porcine reproductive and respiratory syndrome and circovirus ...
... the Journal of Food Science , scientists from The ... 85% of the cocoa flavanols were preserved in recipes for ... cakes, antioxidant activity and cocoa flavanols could be largely retained ... The scientists initially saw that 50 to 95% ...
Cached Biology News:First human gene implicated in regulating length of human sleep 2First human gene implicated in regulating length of human sleep 3K-State lab gives researchers the tools to study porcine circovirus associated diseases 2New study shows that cocoa flavanols can be preserved during cooking and baking 2
... Mouse Antibody to Human Cytokeratin TYPE II ... (Type II) keratins. Including epidermal cell keratin ... 58kD, 65kD and 67kD. Broad keratin reactivity ... and cornea. Immunogen: ...
BD BioCoat Fibronectin 150 mm Culture Dishes...
... number is a new product number, created ... showing no availability yet, please order under ... customer service for assistance. ID ... surface area 500 cm 2 D ...
... for maximum packaging efficiencies of cDNA and genomic ... Using a test insert with Novagen's l vector ... x 10^8 pfu/g ligated l control DNA) are ... to use; simply add up to 5 g ...
Biology Products: