Whole blood RNA quick lysate and application
A lysate and fast technology, which is applied in the field of blood RNA extraction, can solve the problems of unfavorable RNA extraction and time-consuming, and achieve the effect of shortened extraction time, high purity and good integrity
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Embodiment 1
[0036] A rapid whole blood RNA lysate preparation method, each 1000ml volume is composed of the following component mass / volume:
[0037] Guanidine Thiocyanate, 236.32g
[0038] Ammonium thiocyanate, 60.89g
[0039] Anhydrous sodium acetate, 4.102g
[0040] Glacial acetic acid, 14.25ml
[0041] Sodium Lauroyl Sarcosinate, 2.5g
[0042] Sodium chloride, 11.68g
[0043] Glycerin, 75ml
[0044] Water saturated phenol solution, 500ml
[0045] The rest is DEPC treated water.
[0046] Rapidly lyse and extract RNA from 4 parts of 500 μl human anticoagulated whole blood, including the following steps:
[0047] Step 1, Whole blood lysis: add 1ml of the rapid lysis solution to a 2ml centrifuge tube, then add 500μl fresh human anticoagulated whole blood, cover the tube cap, vortex for 30 seconds, and centrifuge at 13,000rpm for 10 minutes; divide into 4 parts Layers are in the same situation, take a photo, such as figure 1 Shown in left 1;
[0048] Step 2: Pass the RNA supernat...
Embodiment 2
[0056] A method for preparing whole blood RNA fast lysate, each 1000ml volume is composed of the following component mass / volume:
[0057] Guanidine Thiocyanate, 118.16g
[0058] Ammonium thiocyanate, 30.45g
[0059] Anhydrous sodium acetate, 8.20g
[0060] Glacial acetic acid, 21.5ml
[0061] Sodium Lauroyl Sarcosinate, 5g
[0062] Sodium chloride, 5.84g
[0063] Glycerin, 50ml
[0064] Water saturated phenol solution, 300ml
[0065] The rest is DEPC treated water.
[0066] A 500μl portion of human anticoagulated whole blood was quickly lysed and RNA was extracted, including the following steps:
[0067] Step 1, Whole blood lysis: Add 1ml of the rapid lysis solution to a 2ml centrifuge tube, then add 500μl fresh human anticoagulated whole blood, cover the tube cap, vortex for 30 seconds, and centrifuge at 13,000rpm for 10 minutes; Such as figure 1 Shown on the right 1;
[0068] Step 2: Pass the RNA supernatant through the purification column: add 400 μl absolute eth...
Embodiment 3
[0073] A method for preparing whole blood RNA fast lysate, each 1000ml volume is composed of the following component mass / volume:
[0074] Guanidine Thiocyanate, 236.32g
[0075] Ammonium thiocyanate, 152.24g
[0076] Anhydrous sodium acetate, 2.75g
[0077] Glacial acetic acid, 9.5ml
[0078] Sodium Lauroyl Sarcosinate, 2.5g
[0079] Sodium chloride, 5.84g
[0080] Glycerin, 75ml
[0081] Water saturated phenol solution, 500ml
[0082] The rest is DEPC treated water.
[0083] RNA extraction was performed on 3 parts of human anticoagulated whole blood stored at -80°C for one month, including the following steps:
[0084] Step 1, whole blood lysis: add 1ml of the rapid lysate to a 2ml centrifuge tube, then add 500μl of thawed human anticoagulated whole blood, cover the tube cap, vortex for 30 seconds, and centrifuge at 13,000rpm for 10 minutes;
[0085] Step 2: Pass the RNA supernatant through the purification column: add 400 μl absolute ethanol to a new 1.5ml centrifug...
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