Gene Keeper RNA & DNA stabilization solution
Code No. 319-08901
Manual Ver. 1 (ND1912-en2401)

Introductions:Gene Keeper RNA & DNA Stabilization Solution (“Gene Keeper”) is apreservation solution for samples used in nucleic acid purification. GeneKeeper rapidly permeates collected samples such as tissues or cells andpreserves nucleic acid integrity. Therefore, the samples can be stably stored
until RNA or DNA purification.

Features.
-In Gene Keeper, samples can be stored for up to 2 days at 37°C, 2 weeks
at 25°C, 1 month at 4°C, and 6 months at -20°C or -80°C.

-

Gene Keeper at -20°C does not freeze easily and does not crystalize easily.

-Samples removed from Gene Keeper can be used for RNA or DNA
purification without washing or wiping.

-

Gene Keeper is suitable for transporting, dividing, and weighing samples
stably without dry ice or liquid nitrogen.

-Contains no toxic substances.

Kit Contents:
Component

Quantity

Storage

Gene Keeper

100 ml x 1

Room temp.** For long-term storage, store the Gene Keeper at 4°C. Gene Keeper are stablefor up to 12 months under refrigeration.

Required Materials Not Included.
-Storage containers or tubes-Tweezers, scissors, scalpel, or needle-Pipettors and pipette tips-

Electronic balance, Centrifuge for cell pellet, if necessary

1Protocol:
Cut samples into slice less than 5 mm thick and submerge in 10 volumes ofGene Keeper. Incubate the sample at room temperature for 1 hour,occasionally mixing by inversion. The sample can be stored in Gene Keeper
for up to 6 months at -20°C or -80°C.
After storage, remove the sample from Gene Keeper with tweezers andproceed to the RNA or DNA purification without washing or wiping.

Procedure: Harvested Animal Tissues (~100 mg)
1. Place 0.5-1 ml of Gene Keeper in a clean storage container on an electronicbalance and tare it. Record its weight.
2. Immediately cut fresh tissue samples to a maximum thickness of 5 mm in any
one dimension, allowing reagent to permeate more easily in the next step.
Alternatively, make notches or puncture with a needle.
(Note) Please try a pilot run on a test sample first to determine the appropriate
procedure treatment depending on your sample type.
(Note) Once a biological sample is harvested, its RNA becomes extremely
unstable until the tissue is completely submerged in a sufficient volume of GeneKeeper. The procedure for tissue harvesting and RNA stabilization should becarried out as quickly as possible.
3. Submerge the sample into the Gene Keeper, weigh the tissue again. If thevolume of Gene Keeper is less than 10 times of the sample weight, add more
Geen Keeper. The minimum volume of Gene Keeper required varies dependingon the sample type, so start with 10 times the amount of the sample.
(Note) Please try a pilot run on a test sample first to determine the appropriate
volume of Gene Keeper depending on your sample type. Larger volumes can beused in necessary. Smaller volumes may lead to RNA degradation during storage.
4. Incubate the sample at room temperature for 1 hour. During incubation, gently
mix by inversion every 15 minutes because the Geen Keeper around the sample
is diluted by the released water from the sample.
(Note) Do not freeze samples in Gene Keeper immediately; store at room
temperature for 1 hour to allow the solution to thoroughly penetrate the tissue.
2(Note) Animal tissue that has been stored in Gene Keeper can be removed fromthe solution, diced into smaller pieces, and returned to Gene Keeper if desired.
5. Store the sample in Gene Keeper at the appropriate temperature. The sample
can be stored in Gene Keeper for up to 2 days at 37°C, 2 weeks at 25°C, 1 monthat 4°C, and at least 6 months at -20°C or -80°C.
(Note) Lower temperature is recommended for longer storage.
6. Remove the sample from Gene Keeper with tweezers and proceed to the RNAor DNA purification. If necessary, dilute the Gene Keeper by adding an equalvolume or more of water to reduce viscosity of the solution, then remove thesample from the solution. There is no need to wash or wipe the sample.
(Note) When extracting RNA using a single reagent, e.g., ISOGEN, make surethat the Gene Keeper content is less than 5% of the final reagent volume (notincluding sample volume).

Procedure: Cultured Cells (~106)
1. Pellet cells by centrifugation according to the protocols followed by yourlaboratory. If necessary, the cells can be washed with PBS.
(Note) Please try a pilot run on a test sample first to determine the appropriate
procedure treatment depending on your sample type.
2. Remove supernatant and add 0.7-1 ml of Gene Keeper into up to 1 x 106 cells.
Immediately suspend the pellet in the Gene Keeper.
(Note) Please try a pilot run on a test sample first to determine the appropriate
volume of Gene Keeper depending on your sample type. Larger volumes can beused in necessary. Smaller volumes may lead to RNA degradation during storage.
3. Store the sample in Gene Keeper at the appropriate temperature. The sample
can be stored in Gene Keeper for up to 2 days at 37°C, 2 weeks at 25°C, 1 monthat 4°C, and at least 6 months at -20°C or -80°C.
(Note) If stored frozen, store at room temperature for a few minutes to allow thesolution to thoroughly penetrate the cells, then store the sample in freezer.
(Note) Lower temperature is recommended for longer storage.

34. Resuspend the cells stored in Gene Keeper, transfer the required amount into
a separate tube. Dilute the Gene Keeper by adding an equal volume or more ofwater to reduce the density of the solution, then centrifuge to pellet cells andremove the supernatant.
(Note) Be sure to remove the cells from Gene Keeper prior to disruption andhomogenization in the RNA or DNA purification procedure.
(Note) When extracting RNA using a single reagent, e.g., ISOGEN, make surethat the Gene Keeper content is less than 5% of the final reagent volume (notincluding sample volume).

Companion Products:
Tissue stored in Geen Keeper is compatible with most RNA/DNA isolation
methods, including single reagent isolation products like ISOGEN and column
purification products like ISOSPIN series.
RNA Purification Kit

Code No.

SampleISOGEN II

311-07361

Tissue, Cell

ISOGEN

311-02501

Tissue, Cell

ISOGEN-LS

311-02621

Liquid sample

ISOSPIN Cell & Tissue RNA

314-08211

Tissue, CellISOSPIN Plant RNA

310-08171

Plant

DNA Purification Kit

Code No.

SampleISOSPIN Tissue DNA

311-07361

TissueISOSPIN Blood & Plasma DNA

312-08131

Blood, Plasma

ISOSPIN Plant DNA

310-08171

Plant

For Research Use Only. Not for diagnostic procedures.
NIPPON GENE CO., LTD.
Head office: 1-5, Kanda Nishikicho, Chiyoda-ku, Tokyo 101-0054 Japan
Laboratory: 2-7-18, Toiya-machi, Toyama 930-0834 Japan
www.nippongene.com
Gene Keeper Manual Ver.1 (ND1912-en2401)

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