Introduction

This catalog contains data for 2066 (AC)n microsatellite markers,
spanning the human genome which includes the previous 1992 catalogue.
Chromosome maps will be made available upon acceptation for publication
of the paper presenting this data.
 
The number of alleles, the heterozygosity and the maximum and minimum
allele sizes (size range) have been determined by observation of 28
unrelated invividuals, namely the grand-parents or the parents of the
CEPH families 1331, 1362, 102, 1347, 1332, 1416, 1413 and 884.

The allele sizes indicated in the reference allele column are those
observed in the the mother of the CEPH family 1347 (individual 134702).
Therefore, 134702 can be used as a reference for allele size
determination. An asterisk indicates markers for which data on 134702
allele size was not available ; in such cases, the size was derived
from the cloned sequence.

PCR amplifications.

All the microsatellite markers described were studied using the
following standard PCR conditions. These conditions represent a
compromise which is probably remote from optimum for most primer
pairs.
PCR reactions are performed in a total volume of 50 µl, containing 40
ng of genomic DNA, 50 pmol of each primer, 0.125 mM dNTPs and 1 unit of
Taq polymerase. 1X amplification buffer contains 10 mM Tris base pH9,
50 mM KCl, 1.5 mM MgCl2, 0.1% Triton X100 and 0.01% gelatin. The
reactions are performed using a "hot-start" procedure: Taq polymerase
is added only after a first denaturation step of 5 minutes at 96"C.
Amplification is carried out during 35 cycles of denaturation (94"C for
40 sec) and annealing (55"C for 30 sec). An elongation step (72"C for 2
min) ends the process after the last annealing.
This "hot-start" procedure permits an efficient primary denaturation
step, and minimizes enzyme degradation. Moreover, addition of Taq
polymerase at a high temperature, considerably reduces mispriming.
Since the amplification products to be obtained are short (90 to 350
base pairs long) and the time interval to raise the temperature from 55
to 94"C (obtained with a ramping rate of 1"C/second) is long enough,
completion of DNA elongation can be achieved without a step at 72"C.
This speeds up the overall amplification.

 

References :
"The 1993-1994 Genethon human genetic linkage map"
Nature Genetics (in press)
Gyapay Gabor, Morissette Jean, Vignal Alain, Dib Colette, Fizames Cecile, 
Millasseau Philippe, Marc Sophie, Bernardi Georgio, Lathrop Marc,
Weissenbach Jean

Abstract :
A genetic linkage map of the human genome has been constructed using
2066 (AC)n short tandem repeats, 60% of which show a heterozygosity of
over 0.7. The map is based on the genotyping of 8 large CEPH families.
Statistical linkage analysis placed these markers in the 23 linkage
groups. The map includes 1266 intervals and spans a total distance of
3690 cM. A total of 1041 markers could be ordered with odds ratios
above 1000:1. About 56% of this map is at a distance of 1 cM or less
from one of its markers.