I am conducting multiplex PCR experiments for infertility detection. And I want to collect DNA from blood sample. However, I do not what the best method for DNA extraction is. What is your experience?
In our lab we use 'QIAamp DNA Blood Mini Kit' from Qiagen; this kit provides quite good DNA yield. We have also compared 2 Qiagen kits, this one and the "DNeasy Blood & Tissue Kit", and DNA quality and yield were better when used QIAamp DNA Blood Mini Kit.
I use Vivantis GF-1 kits. I am satisfied for 2 reasons: 1. It isolate pure DNA without using RNase and Protease K. 2. Its very cheap among other marks.
Www.protocolonline.com have many protcols you can choose the suitable one according your available materials
Qiagen is good but too expensive. The yield with Qiagen can reach using Vivantis GF-1 cheaper
Thanks for all of your comments. I am very confused to choose a kit for this experiment. However, can you give me some problems which you faced when you used the kits and your experience about this troubles? Thanks.
Hi, Dear Thang
I have tried to use a few commercial kits. Some of them were mentioned by our friends, above. I have already used manually methods (alcohol percipitation) . it depends on aim of your study. what do you want to do? how much DNA you need?which post PCR method(s) you want to do? for most common post PCR methods such as RFLP a small yield of of DNA is suffice.
From all kits that I have already tested the best one, in my opinion, is InnuPrep Blood. if you want to extract DNA quickly with high accuracy I would like to introduce this kit. Albeit, I am not the representative of the company:).
I have also attached the manual. you may like to read before using.
Good Luck
I recommend DNA Blood Maxi kit ( #51192), It's easy to use. From 3 ml of Blood recupered on EDTA tube you can have about 50 to 100µg.
Good luck
The selection of your kit depends on three considerations: (1) downstream applications for your DNA; (2) amount you wish to obtain; (3) price per sample that you want to pay.
Just for PCR, most genomic DNA extraction protocols will be OK, from those providing relatively low-quality DNA (such as salting-out procedure) to really high-quality methods. For more stringent applications, such a Southern blotting, MLPA, microarrays or SNP arrays or next-generation sequencing, you must go for the best procedures, providing highest quality unbroken DNA.
In my experience, the very best solution (the gold standard) for genomic DNA extraction from blood is using a Chemagen automated DNA extractor (Chemagic MSM-I from blood). It is state-of-the-art quality for all the most stringent protocols, including NGS. Some research institutes provide this service for a very small per-sample fee. Qiagen manual or automated protocols and Nucleon from GE Healthcare are a good choice too. And you can always consider using DNA Genotek's DNA from saliva technology. It works very well.
The most critical factors regarding template DNA are contaminants such as porphyrin derived from the heme group (if your blood sample was partly hemolyzed or improperly mixed with the antocoagulant) or broken DNA (due to careless manipulation or freezing-thawing cycles). Please keep in mind that genomic DNA must be always stored at 4ºC to avoid double strand breaks caused by freezing.
you can also use the different kits for DNA isolation from blood but Phenol Chloroform method mention in Molecular cloning by Sambrook and Russell is also a good method for DNA isolation from blood in good and pure amount of DNA
In our lab we use 'QIAamp DNA Blood Mini Kit' from Qiagen; this kit provides quite good DNA yield. We have also compared 2 Qiagen kits, this one and the "DNeasy Blood & Tissue Kit", and DNA quality and yield were better when used QIAamp DNA Blood Mini Kit.
Our lab uses the QIAamp DNA Blood Mini Kit and it has been working fine. However, it sort of depends on your downstream processes, I suppose.
https://zageno.com/p/qiaamp-dna-blood-midi-kit/tYk76jp7dXb6YyWfG?q=QIAamp%2520DNA%2520Blood%2520Mini%2520Kit&fs=1
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