By Arun Kumar Sharma, Archana Sharma
Chromosome portray is the main glossy and novel method for at once choosing numerous gene sequences at the same time within the chromosome, using particular probes in molecular hybridization. Its answer levels from unmarried replica to complete genome sequences. it really is now utilized in plant, animal, and human platforms, in gene mapping, id of genetic issues, evolutionary reports, and gene move experiments. This treatise is the 1st of its sort to hide the method with all its changes and purposes. it's designed for normal use by means of postgraduate scholars and study employees in mobile and molecular genetics, plant and animal sciences, agriculture, medication, and phylogenetic studies.
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Extra resources for Chromosome Painting: Principles, Strategies and Scope
2 1 . 04 . Tissue culture flasks with vent cap, 3103-025, Iwaki Ltd . I 3. 1. 05 mM Lglutamine (Iwaki , MED-008), 20 % inactivated fetal bovine serum (FBS; Gibco BRL), 20 ug/ml phytohemagglutinin (HA 15; Murex ), 50 u/rnl penicillin and 50 ug/ml streptomycin . Conduct the culture usin g a mi xture of 9 ml of the culture med ium and I ml of whole blood in a 25-cm 2 tissue culture flask with ve nt cap (lwaki ). Incubate the fla sk in a 5% CO 2 incubator at 37 DC for 70 h. Treat the cultured cells with 50 ng/ml colcemid (G ibco BRL) for 30 min before the harvest.
G ibco - L ife Tech nolog ies. 9800 Me dical Center Drive. PO Box 648 2. Rock ville. MD . USA 06 . Sigma Chemical Co .. O. Box 14508. SI. Lo ui s. 63178 MO. USA 07 . Serva Feinbiochemica. 7 Carl -Benz Strasse, D-6900 Hei del berg. Germa ny 08. Semper. 105 46 Stockholm. Sweden References 01. Heim S. Mite lman F (1995 ). Ca ncer cytogenet ics. 2nd ed . New York: Wiley-Li ss. 02 . Dunham I. Sh imizu N. Roe BA. et aI. (2000). Th e DNA seq ue nce of hum an chromosome 22 . ature 402 : 489-495. 03 . Geno me Directory (1995).
Imag e analvsis Observe the chro mosomes under an Axiophot epiflouresce nce microscope (Carl Ze iss) mounted with a coo led CCD came ra and an auto-wheel filter set regulated by the associated software (PXL ca mera sys tem. Photometries) , Record the data in a computer (Maci ntosh Power PC 8500/120. Apple ) via a scie ntific imagin g software (IPLab Spectrum . Sign al Analytic Corporati on). Save the PRI NS results as a compos ite constructed from the th ree different fl uoresce nce images: DAPI.