Most of Illumina library prep protocols which uses AMPure beads for clean up or size selection recommends using freshly prepared 70% or 80% Ethanol for wash step. Both works very well and I have not noticed any difference in performance of eluted libraries washed by either. I wonder how often or how fresh any one else prepares their wash Ethanol. Fresh prep in literature is justified by presumed or real changes in ethanol concentration by absorption of air moisture. Considering that two recommended wash solutions differs by 10% in Ethanol content does freshness really makes any difference.
Seqanswers Leaderboard Ad
Collapse
Announcement
Collapse
No announcement yet.
X
-
I have recently spoken to a few technicians and post doc about this. Everyone I know makes fresh ethanol for AMPure bead clean ups or size selection - some make 73%, others 74%, 75%, etc. (exact number seems to just be personal preference).
Found this, supporting what Chipper said: "The original AMPure protocol recommended use of a 70% ethanol wash solution. This needed to be freshly made, as over time it becomes more dilute since it absorbs atmospheric water and ethanol evaporates. When you rinse with more dilute ethanol more DNA gets washed away. We have found that this can be avoided by using an 80% ethanol solution instead. This washes just as well but does not need to be freshly prepared each time."
Source
Comment
Latest Articles
Collapse
-
by seqadmin
Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...-
Channel: Articles
04-04-2024, 04:25 PM -
-
by seqadmin
Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...-
Channel: Articles
03-22-2024, 06:39 AM -
ad_right_rmr
Collapse
News
Collapse
Topics | Statistics | Last Post | ||
---|---|---|---|---|
Started by seqadmin, 04-11-2024, 12:08 PM
|
0 responses
25 views
0 likes
|
Last Post
by seqadmin
04-11-2024, 12:08 PM
|
||
Started by seqadmin, 04-10-2024, 10:19 PM
|
0 responses
28 views
0 likes
|
Last Post
by seqadmin
04-10-2024, 10:19 PM
|
||
Started by seqadmin, 04-10-2024, 09:21 AM
|
0 responses
24 views
0 likes
|
Last Post
by seqadmin
04-10-2024, 09:21 AM
|
||
Started by seqadmin, 04-04-2024, 09:00 AM
|
0 responses
52 views
0 likes
|
Last Post
by seqadmin
04-04-2024, 09:00 AM
|
Comment