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Questions related to Nucleic Acid Isolation
Dear Colleagues, I am trying to extract pathogen DNA in a matrix that mimics human sputum. I realize actual clinical samples are best in this regard but my lab at this moment cannot procure any....
19 April 2018 6,348 3 View
I need to purify some genomic DNA from a human skin punch biopsy. In the past, I have done this with the Qiagen DNEasy Blood and Tissue kit. This yields good quality DNA for routine PCRs,...
24 July 2017 4,556 3 View
I am working with Sera-Mag paramagentic beads to isolate whole genomic DNA from human fibroblasts. I follow the general protocol: bind DNA to beads with PEG/NaCl solution, place on magnetic rack,...
23 July 2017 2,990 8 View
I see in the Qiagen handbook that it recommends you buy their proprietary centrifuge in order to get high enough rpms (3000 & 6000) necessary for optimal DNA extractions. I have access to a...
12 July 2017 6,464 2 View
I want to avoid ethanol as it is known to hinder downstream amplification process. Also, I am in a process of developing an automated diagnosis of ZIKA and would want to avoid...
10 July 2017 4,960 3 View
Hi everyone, We have isolated total RNA from our samples using Qiagen RNeasy Plus Micro Kit (which includes genomic DNA removal columns), obtaining RNA yields of around 100 ng. These samples are...
07 July 2017 7,029 3 View
I have approximately 5 x 104 cells after flow sorting for DNA extraction (using many different Qiagen kits), I then always yielded the low quality of DNA for NGS work. Could you please comment on...
07 July 2017 5,861 2 View
Hello everyone, I have been using trizol to extract RNA from small number of cells(10^3), and I added glycogen to facilitate precipitation. What puzzles me is that my control group which...
07 July 2017 3,210 5 View
Hi everyone I'm looking for a DNA extraction protocol from beetles, actually I started extracting DNA from different families (scarabidae, carabidae, cerambycidae) about 2 months ago but there...
28 June 2017 943 14 View
Does anybody have experience with isolation of RNA from a small sample? I am trying to magnetically separate CD4 cells from whole blood and thereafter isolate total RNA from that small cell...
27 June 2017 3,615 3 View
I am trying to extract fungal DNA from clinical samples by using Guanidine thiocynate- phenol saturated . I got good DNA quantity but poor 260/230 ratio. Whether repeatation of CI step can...
10 June 2017 2,048 2 View
Google has failed me. Does anyone have typical (better bad-good-excellent) numbers for the percent recovery of DNA from an ethanol and/or isopropanol precipitation? If it matters, I am using...
06 June 2017 10,041 1 View
I'm looking for a protocol how to isolate gDNA from pet fur/hair obtained after brushing. I know it will only contain very few nucleated cells. Anyone? Preferably without using...
06 June 2017 5,906 2 View
Hi all, I am trying to extract DNA from Alternaria spores. They prove to be very tough to open. Does anyone have a lab protocol they are using to obtain a high yield of Alternaria spore DNA?...
06 June 2017 4,721 25 View
I used Trizol for isolation of RNA from plasma.the 260\230 ratio is very low. how can I solve this problem?
05 May 2017 1,274 3 View
Help! I was performing a phenol purification for phenol: chloroform: IAA solution, and on what I think was my last addition of TRIS, my entire solution turned clear with no visible layering. What...
05 May 2017 7,951 3 View
Hello everyone. I'll be doing a PCR products clean up and I received the suggestion of using AMPure XP since it's easy to distribute among the wells. I'd like to know if someone knows about a...
28 April 2017 1,014 3 View
Dear All, I am looking for a protocol optimized for the DNA (RNA-free) isolation of mammalian (preferentially mouse) MII oocytes. I have different pools of 20-80 oocytes and would like to proceed...
05 April 2017 2,284 1 View
Hi I am hoping to extract RNA from filter samples that have been snap frozen and been placed in -80 since. Previously I have used Trireagent plus Qiagen RNeasy kit as usual and it gives good...
04 April 2017 1,139 1 View
I purchased an Agilent DNA 1000 kit to use in the Bioanalyzer, but I messed up with one spin filter. So, I wanted to filtrate 1 matrix vial with another column already used but it has residual...
08 March 2017 2,389 2 View
For example, EtOH, freezing, RNAlater, etc. The goal is to maximize the amount of high molecular weight DNA for long read sequencing. Thanks!
05 March 2017 6,431 3 View
I have several extraction to perform at the same time and I liked to freeze the yeasts to be able to distribute my extratcions well.
16 February 2017 7,386 3 View
A method that allows me to lyse the trophosites of giardia to obtain the highest yield in DNA extraction.
15 February 2017 2,821 2 View
Please share protocols for faster and efficient lysis of RBC. Probable requirements: lysis buffer to sample (volume) ratio 1:4/1:2
20 January 2017 2,380 9 View