Method for improving electrophoresis resolution capacity and sequencing quality based on nano particle
A nanoparticle and resolution technology, applied in measurement devices, biochemical equipment and methods, material analysis by electromagnetic means, etc., can solve the problems of complex preparation process, inability to adjust and change, etc. The effect of lowering the price of sequencing
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Embodiment 1
[0019] 1) Preparation of sequencing templates and sequencing reactions and other preparations before loading samples according to conventional methods;
[0020] 2) Add 8 μL of loading buffer to each sample, respectively add colloidal gold solution (Sigma Company) with a final concentration of 0.34 nmol / L and a particle size of 10 nm, seal with a common membrane, centrifuge, and mix the loading buffer and colloidal gold The solution is thrown to the bottom of the tube, and the sample in the sample hole is a mixture of the sample to be sequenced, sample buffer and colloidal gold. The samples were then placed into a MegaBACE DNA sequencer for sequencing.
[0021] 3) Use the sequencing software to output the sequencing map and evaluate the effective sequencing length and sequencing quality. As shown in Figure 1, the effective sequencing length of the sample added with colloidal gold is 740nt (the read length of the sequencing map given by the sequencer is 755nt), The sequencing q...
Embodiment 2
[0025] 1) Preparation of sequencing templates and sequencing reactions and other preparations before loading samples according to conventional methods;
[0026] 2) Add 8 μL of loading buffer to each sample, respectively add colloidal gold solution (Sigma Company) with a final concentration of 0.26nmol / L and a particle size of 20nm, seal with a common membrane, centrifuge, and mix the loading buffer and colloidal gold The solution is thrown to the bottom of the tube, and the sample in the sample hole is a mixture of the sample to be sequenced, sample buffer and colloidal gold. The samples were then placed into a MegaBACE DNA sequencer for sequencing.
[0027] 3) The sequencing map was output by sequencing software, and the effective sequencing length and sequencing quality were scored and evaluated. The results showed that the effective sequencing length was increased and the quality score was significantly improved when colloidal gold was added to the loading sample compared w...
Embodiment 3
[0031] 1) Preparation of sequencing templates and sequencing reactions and other preparations before loading samples according to conventional methods;
[0032] 2) Add 8 μL of loading buffer to each sample, respectively add colloidal gold solution (Sigma Company) with a final concentration of 0.02nmol / L and a particle size of 30nm, seal with a common membrane, centrifuge, and mix the loading buffer and colloidal gold The solution is thrown to the bottom of the tube, and the sample in the sample hole is a mixture of the sample to be sequenced, sample buffer and colloidal gold. The samples were then placed into a MegaBACE DNA sequencer for sequencing.
[0033] 3) The sequencing map was output by sequencing software, and the effective sequencing length and sequencing quality were scored and evaluated. The results showed that the effective sequencing length was increased and the quality score was significantly improved when colloidal gold was added to the loading sample compared w...
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