GDP433-RNAprep Pure Blood Kit

RNAprep Pure Blood Kit
For purification of total RNA from human whole blood

RNAprep Pure Blood Kit
(Spin Column)
Cat. no. GDP433

Storage
RNase-Free DNase I, Buffer RDD and RNase-Free ddH2
O (Tubular) should be stored at 2-8°C for 15 months; Buffer RL/β-mercaptoethanol mix can be
stored at 2-8°C for 1 month; others stored at room temperature (15-30°C)
for 15 months.

Introduction
RNAprep pure Blood Kit provides a fast, simple, and cost-effective method
for purification of total RNA from blood. The purified RNA is ready for use
in downstream applications such as RT-PCR and real-time RT-PCR, micro
array, northern blot, dot blot, polyA screening, in vitro transcription, and
molecular cloning.

Notes of preventing RNase contamination

1. Change gloves regularly. Bacteria on the skin can result in RNase
   contamination.
2. Use RNase-Free plastic and tips to avoid cross-contamination.
3. RNA can be protected in Buffer RL, but RNA must be stored or applied
   in RNase-Free plastic or glassware. To wipe off RNase, the glassware can
   be roasted at 150°C for 4 hours, while plastic can be dipped in 0.5 M
   NaOH for 10 min, washed by water thoroughly, then sterilization.
4. Use RNase-Free water to confect solution.

Important points before starting

1. β-Mercaptoethanol (β-ME) must be added to Buffer RL before use. The
   final concentration of β-ME is 1%. For example, add 10 μl β-ME to 1 ml
   Buffer RL. Buffer RL containing β-ME can be stored at 2-8°C for up to 1
   month. Buffer RL may form a precipitate upon storage. If necessary, redissolve by warming, and then place at room temperature (15-30°C).
2. Buffer RW is supplied as a concentrate. Before using for the first time,
   add ethanol (96-100%) as indicated on the bottle to obtain a working
   solution.
3. Blood and body fluids of all human subjects are considered potentially
   infectious. All necessary precautions should be taken when working
   with whole blood.
4. The maximum amount of human whole blood that can be processed (1.5
   ml) has been determined for blood from healthy adults (approximately
   4000-7000 leukocytes per microliter). Reduce amount appropriately if
   using blood with elevated numbers of leukocytes. A maximum of 1 ×10
   7leukocytes can be processed.
5. After leukocyte lysis, all steps of this protocol should be performed at
   room temperature (15-30°C), as quickly as possible.
6. Homogenized cell lysates (in Buffer RL) can be stored at -70°C. Frozen
   lysates should be incubated at 37°C in a water bath for 10 min until
   completely thawed and salts are dissolved. Continue with step 7.
7. Frozen whole blood cannot be used.

Preparation of DNase I stock solution
Dissolve the lyophilized DNase I (1500 U) in 550 μl of the RNase-Free
ddH2O (Tubular). Mix gently by inverting. Do not vortex. Divide it into
single-use aliquots, and store at -30~-15°C for up to 9 months.
Note: Thawed aliquots can be stored at 2-8°C for up to 6 weeks. Do not
refreeze the aliquots after thawing.

GDP433-RNAprep Pure Blood Kit

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