Custom DNA Sequencing:Sample Preparation | Hokkaido System Science Co.,Ltd.

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Sample Preparation

Regarding the sample preparation

	When ordering a DNA sequencing, please accompany an agarose gel electrophoresis image to the sample. It is used to verify the status and concentration of the sample. Please follow the guidelines below for getting a better result.
About the sample
	Please use sterile deionized water rather than buffer with EDTA when preparing the sample and the primer since EDTA may affect the cycle sequencing reaction adversely.

About concentration of the sample

Spectrophotometry is not suitable for measuring concentration of a sample to be subject to sequencing. Please measure concentration with electrophoresis, i.e., by comparison to a molecular mass marker. (Reading may differ by, as much as 10 folds.)
Please take a electrophoresis using 1 µl of apply as a guide. The image to accompany the sample should show the band clearly.
Please make sure the electrophoresis image looks like the samples.

For a plasmid sample

For a PCR product


	Shipping volumes

For a plasmid sample (volume per sequencing reaction)

Sample size	Necessary concentration	Necessary volume
2-4 kbp	0.2 µg/µl=200 ng/µl	10 µl or more
6 kbp	0.3 µg/µl=300 ng/µl	10 µl or more
8 kbp	0.4 µg/µl=400 ng/µl	10 µl or more
10 kbp	0.5 µg/µl=500 ng/µl	10µl or more

For a PCR product (volume per sequencing reaction)

Sample size	Necessary concentration	Necessary volume
100 – 500 bp	10 ng/µl	10 µl or more
500 – 1000 bp	10 - 20 ng/µl	10 µl or more
1000 – 2000 bp	20 - 40 ng/µl	10µl or more
> 2000 bp	40 - 100 ng/µl	10 µl or more

Primer

Please send 10 µl or more of primer of approximately 10 µM per reaction.