Hello,

Home-made recipe for 1 L:

Reagents                                                                   Final Concentration

Phenol in saturated buffer 380 ml                            38 %

Guanidine thiocyanate 94.53 g                                 0.8 M

Ammonium thiocyanate 76.12 g                               0.4 M

Sodium acetate, pH 5.0 33.4 ml of 3 M stock         0.1 M

Glycerol 50 ml                                                              5 %

DEPC-Water Adjust the final volume to 1 L

Hope it helps

Regards

Ai

ni

pH 4.5.    Pure phenol.   10 volumes of Trizol to one volume loose packed (70 x g).  Shear DNA with 25g needle.  Spin 12 x g.  Collect 80% of aqueous phase.

I use for the THP-1 either premade TRIZOL or to isolate the RNA the RNA-easy kit from Qiagen.

Here are some interesting patents that may be of use to you. Please, see them.

1. US5346994

2. US4843155

3. US8367817 B2

Out of curiosity, couldn't you just DNAse treat your final isolates?

@John Hildyard i tried many times but RNA is getting degraded after DNase treatment, i am using Thermoscientific DNase 1

See the classic paper by Saatchi and Chomcinski

Hello,

Home-made recipe for 1 L:

Reagents                                                                   Final Concentration

Phenol in saturated buffer 380 ml                            38 %

Guanidine thiocyanate 94.53 g                                 0.8 M

Ammonium thiocyanate 76.12 g                               0.4 M

Sodium acetate, pH 5.0 33.4 ml of 3 M stock         0.1 M

Glycerol 50 ml                                                              5 %

DEPC-Water Adjust the final volume to 1 L

Hope it helps

Regards