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Transgenic Core
Director: Arlene Sharpe
Location: 77 Avenue Louis Pasteur, NRB 837, Boston

 
Core Summary:

Transgenic Core Facility provides services for generating transgenic and knockout mice. Established in 1992, the BWH Core Facility has been providing gene targeting and microinjection services using state-of-the-art facilities and equipment.

Personnel/Contact Information:

Director: Arlene Sharpe, M.D, Ph.D.
Phone: (617) 432-6569
Fax: (617) 432-6570
Email:
asharpe@rics.bwh.harvard.edu

Other Staff:

Transgenic Specialist: Lina Du, M.D.
Phone: (617) 432-6227
Email:
lina_du@hms.harvard.edu

ES Cell Culture Technician: Xiaomeng Tang
Phone: (617) 432-6182
Email:
xtang@rics.bwh.harvard.edu

ES Cell Culture Technician: Matthew Finn
Phone: (617) 432-6576
Email:
matthew_finn@hms.harvard.edu

Facilities and Equipment:

Location of Core:

The facility is located at 77 Avenue Louis Pasteur, NRB 837, Boston.

Major Equipment:

The Core Facility has all of the equipment necessary for generation of transgenic mice including a Nikon Diaphot microscope equipped with Nomarski Optics and Narishige micromanipulators for microinjections, Nikon surgical microscopes for egg isolation ad transfer, a sutter needle puller and a de Fonbrune microforge.

Services:
  • Microinjection of DNA constructs (including BACs) into pronuclei for generating transgenic mice (FVB or C57 BL/6 strain background)
  • Microinjection of embryonic stem (ES) cells into blastocysts for generating knockout mice
  • ES cell transfection for generating ES cell clones with the targeted combination event
  • Lentiviral transgenics
  • Microinjection of IPS
Fee Schedule/Rates:

Service
BWH Investigator *
Non-BWH Investigator

ES Cell Transfection Service (129ES)

$2,600
$3,800

ES Cell Transfection Service (C57BL/6)

$3,000
$4,300
Thawing and Expanding Cells
$625
$850
ES Cell Preparation for Microinjection
$200
$275
DNA Injections (FVB)
$3,750
$4,600
DNA Injections (C57 BL/6)
$4,475
$5,450
Lentiviral Transgenic
$3,750
$4,600
DNA Injections (BALB C)
$5,325
$6,375
BAC Injections
$5,000
$6,100
ES Cell Injections 129 (Blastocyst)
$1,450
$2,200
ES Cell Injections B6 (Blastocyst)
$1,550
$2,300
Inducible Progenitor Stem Cells (IPS) Injections
$1,450
$2,200
DNA Injection and Microdissection
$1,500
$2,250
Additional Injection
$1,500
$2,250
Tail biopsy
$175
$200
Breeding Services
(Per diem/Purchase) Plus $150/mo
(Per diem/Purchase) Plus $200/mo
Technical Services Charge/hr
$75/hr
$100/hr
Exportation Fee (includes shipping supplies and associated shipping costs)
$75
$125
* BWH status determined by institutional source of payment.

Getting Started:

Our ES facility will generate targeted ES cell lines with KO constructs provided by investigators. Upon receipt of the constructs and application form, electroporation will be scheduled. The Core Director, Dr. Arlene Sharpe will prioritize the service requests according to the difficulty of the project and work load.

  1. Transfection of your linearized DNA construct into ES cells, J1 (129/SV) and Bruce 4 (C57 BL/6J).
  2. After electroporation and selection, clones will be picked, cultured in multi-well plates and replicated. The clones will be frozen to be used to expand positive clones.
  3. The investigator will be notified and given DNA isolated from from each clone suitable for screening.
  4. Since storage of frozen cells in multi-well dishes is not optimal, it is strongly suggested that researcher return the screening results to the core within 4 months of receipt of the DNA samples.
  5. After screening the clones, up to six ES positive clones will be expanded and frozen down.
  6. After expansion, the investigator will then receive DNA from each clone for reconfirmation of targeted recombination. After reconfirmation, the best growing clones will then be scheduled for injection into blastocysts.  Chimeric mice will be given to the investigator.
Links to other outside Academic and Commercially Available Services:

Institution Contact Website
Tufts Univeristy Janis Lem
jlem@tufts-nemc.org
http://www.nemc.org
BIDMC Joel Lawitts
(617) 632-0264
http://research.bidmc.harvard.edu/CoreLabs/trans.asp
MIT Aurora Connor
(617) 452-3658
http://web.mit.edu/transgenic/
Johns Hopkins Chip Hawkins
(410) 614-3858
http://www.hopkinsmedicine.org/core/Home.htm

Departmental Web Link:

http://www.bwhtransgenic.org/index.html