Mouse rederivation from frozen embryos (embryos from external stock)
deliverables
price
at least 4 weaned rederived animals (3) or 1 mutant, free of pathogens on Felasa list if embryo transfer is not successful due to - not enough embryos to do a transfer (< 10) - fosters did not get pregnant - no or less then 4 offspring and no mutants - sperm freezing
1100 € (1) (2)
at least 4 weaned rederived animals (3) or 1 mutant, free of pathogens on Felasa list if IVF or embryo transfer is not successful due to - not enough embryos to do a transfer (< 10) - fosters did not get pregnant - no or less then 4 offspring and no mutants at least 4 weaned rederived animals (3) or 1 mutant, free of pathogens on Felasa list if transfer is not successful due to - not enough embryos to do a transfer (< 10) - fosters did not get pregnant - no or less then 4 offspring and no mutants
1320 € (1)
(1)
cost of health screening not included
(2)
2 attempts are performed
(3)
irrespective of genotype
(9)
if sperm needs to be frozen especially for the rederivation proces
550 € (1) (2) 550 € (9)
550 € (1) 1100 (1) 550 (1)
STRAIN RECONSTITUTION Service Mouse sperm revival I (sperm from 'safely frozen' TMCF sperm stock) Mouse sperm revival II (sperm from TMCF sperm stock, but not 'safely frozen')
Mouse embryo revival I (embryos from 'safely frozen' TMCF stock) Mouse embryo revival II (embryos from TMCF stock, but not safely frozen) (1)
cost of health screening not included 2 attempts are performed (3) irrespective of genotype (2)
deliverables at least 4 weaned rederived animals (3) or 1 mutant, free of pathogens on Felasa list
price 1320 € (1)
at least 4 weaned rederived animals (3) or 1 mutant, free of pathogens on Felasa list
1320 € (1) (2)
- not enough embryos to do a transfer (< 10) - fosters did not get pregnant - no or less then 4 offspring
550 € (1) (2)
at least 4 weaned rederived animals (3) or 1 mutant, free of pathogens on Felasa list
1100 € (1)
at least 4 weaned rederived animals (3) or 1 mutant, free of pathogens on Felasa list
1100 € (1) (2)
- fosters did not get pregnant - no or less then 4 offspring
550 € (1) (2)
CRYOPRESERVATION Service
deliverables
price
Sperm cryopreservation: available line sperm freezing + QC = ONLY ON REQUEST + SIGNATURE PI!!
at least 5 straws frozen
1.650 €
Sperm cryopreservation: unique line Safety backup: Stop the breeding: Embryo cryopreservation (6) (4)
QC: IVF > 20% fertilization Sperm freezing only (no QC): at least 5 straws frozen Sperm freezing and QC: at least 5 straws frozen QC IVF > 20% fertilization at least 200 homozygous or 300 heterozygous embryos frozen
free if - strain is euthanized within 3 months after being notified strain is successfully frozen 500 € if strain is still alive within 3 months after being notified strain is successfully frozen (5) (6)
up to 3 freezings will be done if a previous freezing was not successful in the IVF only on special request
550 €
(5)
free free
(4)(5)
case by case (1100-3300€)
ES-CELL TARGETING Service
deliverables
- 24 well DNA from 200-400 picked ES-cell clones - triplicate frozen -80 stock of each clone on 96 well plates - thawing up to 8 positive clones ES-cell targeting I - expanding and freezing stock of 2-3 vials/clone in LN2 (vector integrates by HR) - DNA preparation of each positive clone for reconfirmation - karyotyping of each positive clone by chromosome count - 1 new electroporation in case 1st electroporation didn't result in positive clones - 24 well DNA from 10 picked ES-cell clones - triplicate frozen -80 stock of each clone on 96 well plates - thawing up to 8 positive clones ES-cell targeting II - expanding and freezing stock of 2-3 vials/clone in LN2 (vector integrates by RMCE) - DNA preparation of each positive clone for reconfirmation - karyotyping of positive clones by chromosome count - 1 new electroporation in case 1st electroporation didn't result in positive clones - 24 well DNA from 100 picked ES-cell clones - triplicate frozen -80 stock of each clone on 96 well plates - thawing up to 8 positive clones ES-cell targeting III - expanding and freezing stock of 2-3 vials/clone in LN2 (vector integrates randomly) - DNA preparation of each positive clone for reconfirmation - karyotyping of each positive clone by chromosome count - 1 new electroporation in case 1st electroporation didn't result in positive clones - 24 well DNA from 5-10 picked ES-cell clones sensitive to selection marker that is removed - triplicate frozen -80 stock of each clone on 96 well plates - thawing up to 8 positive clones ES-cell targeting IV (Flpe or Cre electroporation to remove - expanding and freezing stock of 2-3 vials/clone in LN2 selection marker) - DNA preparation of each positive clone for reconfirmation - karyotyping of each positive clone by chromosome count - 1 new electroporation in case 1st electroporation didn't result in positive clones
price
2.750 €
1.650 €
2.200 €
1.100 €
GENERATION OF MUTANT MICE Service generation of chimeras by aggregation or blastocyst injection generation of tg mice by zygote injection of cDNA generation of tg mice by zygote injection of BAC DNA
generation of mutant mice by zygote injection of nucleases I (deletion-mutation using NHEJ)
generation of mutant mice by zygote injection of nucleases II (deletion-mutation using NHEJ)
deliverables - karyotyping clones by chromosome counting (if not done yet for external clones) - aggregation of 160 morulas or injection of 80 blastocysts or 2 high (>50%) chimeras, whatever comes first - testbreeding of chimeras to check for germline transmission - in silico design, cloning, purification and linearisation of cDNA construct - injection of 400 C57BL/6J zygotes or 2 founders, whatever comes first - in silico design, cloning and purification of BAC vector - injection of 400 C57BL/6J zygotes or 2 founders, whatever comes first - in silico design + construction of guideRNA - sgRNA preparation and purification - sequence verification of sgRNA construct - in silico design PCR screening strategy - optimization of PCR screening - in vitro test functionality of sgRNA - injection of 400 C57BL/6J zygotes or 2 founders, whatever comes first - injection of 400 C57BL/6J zygotes or 2 founders, whatever comes first
- design + construction of guideRNA - nuclease RNA preparation generation of mutant mice by zygote injection of nucleases + donor DNA I (deletion- design donor DNA mutation by HDR) - ordering ss or ds donor DNA(10) - injection of 600 C57BL/6J zygotes or 2 founders, whatever comes first generation of mutant mice by zygote injection of nucleases + donor DNA II (knockin - injection of 600 C57BL/6J zygotes or 2 founders, whatever comes first using HDR)
(7)
cage prices for fosters, offspring from fosters, testbreedings, offspring from testbreedings not included depending on size of donor DNA (10) cost of the donor DNA is not included in the end-price and should be payed by the researcher (11) knockin-founders cannot be guaranteed (8)
price 2750 €
(7)
1.100 € 2750 € (7) 1.650 € 2750 € (7)
3300 € (7)
2750 €
(7)
4400 €
(8)(11)
3850 €
(8)(11)
ES-CELL LINE DERIVATION Service
derivation of ES-cell lines
deliverables - flushing blastocysts from uterus - keeping blastocysts in ES-derivation culture medium - picking and expanding outgrowths from blastocysts - karyotyping of each line (up to 10) by chromosome counting - 24 well DNA preparation from each line - freezing 2-3 vials from each line and storing in LN2
