Catalog#

R6617-00

R6617-01

R6617-02

Preps

4

50

250

Buffer LY

2.4 mL

28 mL

135 mL

Buffer RB

3 mL

30 mL

135 mL

RNA Wash Buffer *

2 mL

20 mL

3 x 24 mL

DEPC-Treated ddH₂O

500 µL

10 mL

30 mL

DNase Stop Buffer

200 µL

2.4 mL

12 mL

ezBind Columns

4

50

250

Collection Tubes

8

100

500

Lyticase

13 U

155 U

775 U

User Menu

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R6617-00

Dissolve with 130 μL of Elution Buffer

R6617-01

Dissolve with 1.6 mL of Elution Buffer

R6617-02

Dissolve with 8 mL of Elution Buffer for each bottle

Problem

Possible reason

Suggested Improvement

Low A₂₆₀/A₂₈₀ ratios

Protein contamination

Do a Phenol: Chloroform extraction. Loss of total RNA (up to 40%) should be expected.

Guanidine Thiocyanate contamination

Add 2.5 volumes of ethanol and 0.1M NaCl (final concentration) to precipitate RNA. Incubate for 30 min at -20 ^oC. Centrifuge at 10,000 g for 15 min at 4 ℃. Resuspend the RNA pellet in DEPC-ddH₂O.

Low Yield

RNA in sample degraded

Freeze samples immediately in liquid nitrogen and store at -70 ^oC after collect it.

The binding capacity of the membrane in the spin column was exceeded

Use of too much tissue sample exceeding the binding capacity of spin column will cause the decreasing of total RNA yield.

Ethanol not added to buffer

Add ethanol to the RNA Wash Buffer and DNase Stop Buffer before purification.

Genomic DNA contamination

Too much total RNA sample was used in RT-PCR.

Reduce total RNA amount used in RT-PCR to 50-100 ng.

The sample may contain too much genomic DNA.

Reduce the amount of starting tissue in the preparation of the homogenate. Most tissues will not show a genomic DNA contamination problem at 30 mg or less per prep.

Reduce cell numbers to 1-2x10⁶ or increase buffer volume and do multiple loadings to column.