Author: Monica Sentmanat; Evguenia Kouranova; Xiaoxia Cui
Title: One-step RNA extraction for RT-qPCR detection of 2019-nCoV Document date: 2020_4_5
ID: ihhu7nef_10
Snippet: In the parallel comparison of QE buffer lysed samples versus column purified samples, the Ct values for positive controls and RNaseP were within 1 cycle or less, even though the samples were concentrated 4x (from 120 ul to 30 ul elute). It indicates that there is significant loss of material by column purification and the benefit of concentrating the samples is evened out. Additionally, our method starts with only 200 ul of lysis buffer, and 2 ul.....
Document: In the parallel comparison of QE buffer lysed samples versus column purified samples, the Ct values for positive controls and RNaseP were within 1 cycle or less, even though the samples were concentrated 4x (from 120 ul to 30 ul elute). It indicates that there is significant loss of material by column purification and the benefit of concentrating the samples is evened out. Additionally, our method starts with only 200 ul of lysis buffer, and 2 ul of direct lysis is 1/100 of the total sample. If we use a smaller swab, which is available, the volume can be lowered further. With the standard method, the sample transport buffer is 3 mL and only 120 ul to 140 ul, depending on the extraction kit, is taken for purification, essentially a 1:15 dilution that can translate into up to a difference of 4 Ct's.
Search related documents:
Co phrase search for related documents- lysis buffer and positive control: 1, 2, 3
- lysis buffer and RNaseP positive control: 1
- lysis buffer and sample concentrate: 1
- lysis buffer and significant loss: 1
- lysis buffer and standard method: 1, 2, 3
- lysis buffer and total sample: 1, 2, 3
- lysis buffer and transport buffer: 1, 2
- lysis buffer and volume lower: 1
- positive control and RNaseP positive control: 1, 2, 3
- positive control and significant loss: 1, 2, 3, 4, 5
- positive control and standard method: 1, 2, 3, 4
- positive control and total sample: 1
- sample transport buffer and transport buffer: 1, 2
- significant loss and standard method: 1, 2
- significant loss and volume lower: 1, 2
- small swab and standard method: 1
Co phrase search for related documents, hyperlinks ordered by date