390
ILSE CHUDOBA and GABRIELE SENGER
- Microliter pipettes
Again, due to the high risk of DNA contamination, we reserve one set
of microliter pipettes (0.01-2 Ill, 0.1-20 Ill, 10-200 Ill) for the manipulation prior to the DOP-PCR and for the DOP-PCR itself, respectively.
For all manipulations after the DNA fragments have been tagged with
the DOP-primers, different pipettes are used.
- Needle holder
A reftllable lead pencil is used to hold the needles.
Consumables - Coverslips (24x60 mm)
Due to the arrangement of an inverted microscope and the oil immersion objective, coverslips are used for spreading metaphases instead of
regular microscope glass slides.
- Filter tips (suitable for the microliter pipettes)
For all manipulations of liquids, filter tips are used.
- PCR tubes
Depending on the thermocyder 500- or 200-111 tubes are used.
- Plastic centrifuge tubes, sterile, (50 ml) (Greiner, #227 261)
Reagents - DOP-Primer (=6MW) (Telenius et al. 1992) 5' ccg act cga gn6 atg tgg 3
- Sodium dodecyl sulfate (SDS) (Sigma, L 4509)
- Bacto Trypsin (Difco, #0153-61-1)
- Phosphate-Buffer, pH 6.88 (Merck, #7294)
- Giemsa-solution (Merck, #9204)
- Proteinase K (Boehringer, #1413 783)
- Sequenase, Version 2.0, (Amersham, #US7077S)
- Taq DNA Polymerase (Appligene Oncor, #120186AX)
- dNTP set, PCR grade (Boehringer Mannheim, #1969 068)
- Chromatide Texas Red-12dUTP (Molecular Probes, #C-7631)
- 20xSSC (Eppendorf, #0032 006.701)
- E. coli tRNA (Boehringer, #109 541)
- Deionized Formamide (Appligene Oncor, #S4117)
ILSE CHUDOBA and GABRIELE SENGER
- Microliter pipettes
Again, due to the high risk of DNA contamination, we reserve one set
of microliter pipettes (0.01-2 Ill, 0.1-20 Ill, 10-200 Ill) for the manipulation prior to the DOP-PCR and for the DOP-PCR itself, respectively.
For all manipulations after the DNA fragments have been tagged with
the DOP-primers, different pipettes are used.
- Needle holder
A reftllable lead pencil is used to hold the needles.
Consumables - Coverslips (24x60 mm)
Due to the arrangement of an inverted microscope and the oil immersion objective, coverslips are used for spreading metaphases instead of
regular microscope glass slides.
- Filter tips (suitable for the microliter pipettes)
For all manipulations of liquids, filter tips are used.
- PCR tubes
Depending on the thermocyder 500- or 200-111 tubes are used.
- Plastic centrifuge tubes, sterile, (50 ml) (Greiner, #227 261)
Reagents - DOP-Primer (=6MW) (Telenius et al. 1992) 5' ccg act cga gn6 atg tgg 3
- Sodium dodecyl sulfate (SDS) (Sigma, L 4509)
- Bacto Trypsin (Difco, #0153-61-1)
- Phosphate-Buffer, pH 6.88 (Merck, #7294)
- Giemsa-solution (Merck, #9204)
- Proteinase K (Boehringer, #1413 783)
- Sequenase, Version 2.0, (Amersham, #US7077S)
- Taq DNA Polymerase (Appligene Oncor, #120186AX)
- dNTP set, PCR grade (Boehringer Mannheim, #1969 068)
- Chromatide Texas Red-12dUTP (Molecular Probes, #C-7631)
- 20xSSC (Eppendorf, #0032 006.701)
- E. coli tRNA (Boehringer, #109 541)
- Deionized Formamide (Appligene Oncor, #S4117)