BN QuickPick^TM mRNA
For the rapid purification of mRNA

The methods in the Bio-Nobile QuickPick mRNA kits are based on the binding of poly-A⁽⁺⁾  mRNA on immobilised oligo (dT)₃₀ on paramagnetic particles  in the following steps.

1.    Cells are first lysed in Lysis/Binding Buffer
2.    Cell lysate is mixed with washed oligo (dT)₃₀ coated magnetic particles, incubated for 4 minutes at room temperature for hybridizing      
        oligo (dT) to mRNA.
3.    Particle-bound mRNA is separated from other RNA species, such as pre-mRNA, ribosomal RNA, tRNA, snRNA, by washing.  The particle 
       bound mRNA is resuspended in Elution Buffer, eluted at +70⁰ C for 5 minutes.
       The procedure starting from cells and ending with pure mRNA last 10 minutes.

BN QuickPick^TM mRNA isolation protocol

Volumes vary depending on kit used.
Number tubes 1 to 8 
Tube 1 : Oligo (dT)₃₀ coated well-mixed magnetic particle suspension in storage buffer.
Tube 2 : Lysis/Binding Buffer
Tube 3 : cellular sample in Lysis/Binding Buffer
Tube 4 : Wash Buffer A
Tube 5 : Wash Buffer A
Tube 6 : Wash Buffer B
Tube 7 : Appropriate buffer for downstream application.  Optional (not supplied in kit)
Tube 8 : Elution Buffer

Protocol

1.    Collect magnetic particles from Tube 1with PickPen and wash by releasing then into Tube 2 (Lysing/Binding Buffer).  Mix.

2    Collect magnetic particles from Tube 2 with PickPen and release them into Tube 3 (Sample). Mix gently at room temperature for 4 minutes and allow mRNA to anneal to the magnetic particles.

3.    After annealing collect  the magnetic particles with PickPen from Tube 3 transfer to Tube 4 and wash ( Wash Buffer A).  Repeat the washing step by transferring the particles to Tube 5 (Wash Buffer A)  and  Tube 6 (Wash Buffer B)  and to Tube 7 (appropriate buffer for the downstream application, optional).  Finally release the magnetic particles into Tube 8 (Elution Buffer) and mix thoroughly. mRNA bound to the beads can be eluted from the beads by heating at +700 C for 5 minutes.

4.    After elution collect the beads with the PickPen and discard the magnetic particles.  Transfer the eluate into a suitable tube containing        downstream application buffer.  Mix.  Alternatively, the eluate can be stored at -700 C.

mRNA isolated from human U 937
cells using QuickPick mRNA mini kit

                             Note:  SML Kits are scalable.  PickPen Tips are not included in the Kit.  The number of preparations  
                             quoted have a reference volume of 100 mL          

For details of the PickPen^TM magnetic transfer device click here

FOR RESEARCH USE ONLY - NOT FOR USE IN DIAGNOSTIC PROCEDURES

Metachem Diagnostics Ltd.
29 Forest Road, Piddington, Northampton. NN7 2DA
Tel. (01604) 870370     Fax (01604) 870194