1.0 OBJECTIVE
1.1
To lay down a standard procedure to be followed for separation of serum/plasma from whole
blood for pre-clinical experiments in Animal House Facility.
2.0
SCOPE
2.1
This Standard Operating Procedure (SOP) shall be
applicable for the separation of serum/plasma from whole
blood for pre-clinical experiments in Animal House Facility.
3.0
RESPONSIBILITY
3.1
Study Director / designee
3.2
All personnel involved in the activity of separation of
serum/plasma from whole blood
4.0
DEFINITIONS
4.1
Nil
5.0
PROCEDURE
5.1
Separation of
Serum
5.1.1
Blood shall be collected from animals as per current
running version of SOP…….. into labeled collecting vials without anticoagulant.
5.1.2
Vials shall be labeled properly with a study number,
animal number, Animal Group, sex, etc.
5.1.3
After collection, mix the blood gently. Don't shake the
sample vigorously.
5.1.4
Store the vials at room temperature in an upright
position to allow coagulation. Clot formation should be completed in 20-30 min
in most cases.
5.1.5
Centrifuge to separate the serum from the blood clot
with centrifugation parameters i.e. 2-8 °C, 10-15 min, 4000-5000 RPM (or as per
study plan).
5.1.6
Storage time at room temperature (until centrifugation)
should not exceed 40 min.
5.1.7
Transfer the serum into pre-cooled labeled collection
tubes (e.g. Falcon) without aspirating blood cells. Use disposable pipette
tips. Keep serum on ice.
5.1.8
Labeled sample storage tubes shall be pre-cooled and
serum sample shall be transferred into pre-cooled vials in suitable aliquots to
avoid later freeze/thaw cycles.
5.1.9
Serum samples shall be frozen immediately at -20 °C for
long term use.
5.1.10
Store the vials in boxes, ordered by sample number and
other study-related information.
5.2
Separation of
Plasma
5.2.1
Calculated amount of anticoagulant sodium citrate/ K3EDTA
shall be taken in clean labeled Eppendorf vials.
5.2.2
Blood shall be collected from animals into labeled
collecting vials as per the current running version of SOP………..
5.2.3
Vials shall be labeled properly with a study number,
animal number, Animal Group, sex, etc.
5.2.4
After collection, mix the blood gently. Don't shake the
sample vigorously.
5.2.5
Centrifuge to separate the plasma from the blood with
centrifugation parameters i.e. 2-8 °C, 10-15 min, 4000-5000 RPM (or as per
study plan)
5.2.6
After centrifugation, the plasma layer will be separated on
the top.
5.2.7
Storage time at room temperature (until centrifugation)
should not exceed 40 min.
5.2.8
Plasma layer shall be carefully collected into other
labeled tube with the help of appropriate pipette without disturbing the buffy
coat layer.
5.2.9
Transfer the plasma into pre-cooled labeled collection
tubes (e.g. Falcon) without aspirating blood cells. Use disposable pipette
tips. Place plasma on ice.
5.2.10
Labeled sample storage tubes shall be pre-cooled and
plasma sample shall be transferred into pre-cooled vials in suitable aliquots
to avoid later freeze/thaw cycles.
5.2.11
Plasma samples shall be frozen immediately at -20 °C
for long term use.
5.2.12
Store the vials in boxes, ordered by sample number and
other study-related information.
6.0
ENCLOSURES
6.1
Nil
7.0
ABBREVIATIONS
7.1
SOP : Standard Operating Procedure
7.2
QAD : Quality Assurance Department
7.3
AHF : Animal House Facility
8.0
REFERENCES
8.1
In-house
9.0
REVISION HISTORY
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Sr. No. |
Change Control No. |
Reason for change |
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END OF
DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the In-vivo Laboratory.
4. List of All SOPs and Documents for Clinical Research
5. List of Instruments and Equipment Used In Laboratory Animal Facility
6. List of All SOPs and Documents for the Animal House Facility
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