P103
Homo sapiens
Protocols: Peripheral venous blood was collected from both patients and their families. Buffy coats of nucleated cells obtained from anticoagulated blood (ACD or EDTA) were resuspended in 15 ml polypropylene centrifugation tubes with 3 ml of nuclei lysis buffer (10 mM Tris-HCl, 400 mM NaCl and 2 mM Na2EDTA, pH 8.2). The cell lysates were digested overnight at 37°C with 0.2 ml of 10% SDS and 0.5 ml of a protease K solution (1 mg protease K in 1% SDS and 2 mM Na2EDTA). After digestion was complete, 1 ml of saturated NaCl (approximately 6M) was added to each tube and shaken vigorously for 15 seconds, followed by centrifugation at 2500 rpm for 15 minutes. The precipitated protein pellet was left at the bottom of the tube and the supernatant containing the DNA was transferred to another 15 ml polypropylene tube. Exactly 2 volumes of room temperature absolute ethanol was added and the tubes inverted several times until the DNA precipitated. The precipitated DNA strands were removed with a plastic spatula or pipette and transferred to a 1.5 ml microcentrifuge tube containing 100-200 pl TE buffer (10 mM Tris-HCl, 0.2 mM Na2EDTA, pH 7.5). The DNA was allowed to dissolve 2 hours at 37°C before quantitating. AmpliSeqTM ibrary was prepared following the modifiedIon AmpliSeqTM Library Preparation protocol (Pub No. MAN0006735).
ENA first public
2018-02-02
ENA last update
2018-01-21
External Id
SAMEA104499834
INSDC center alias
Peking University First Hospital
INSDC center name
Peking University First Hospital
INSDC first public
2018-02-02T17:04:51Z
INSDC last update
2018-01-21T22:08:18Z
INSDC status
public
Submitter Id
E-MTAB-6404:P103
age
6
broker name
ArrayExpress
common name
human
disease
hypomyelinating leukodystrophy
organism part
blood
sample name
E-MTAB-6404:P103
sex
female