1) Puncture bacteria
Please use a single toothpick to pierce the solid medium that is supplemented with proper antibiotics. After overnight incubation, the bacteria can be identified and sent.
2) Bacteria grown in liquid media
Growth should be visible to the naked eye and is enriched after overnight incubation. 200ul of the bacteria in growth medium should be added to 1.5ml sterilized centrifuge tubes. The name of the sample should be marked. In addition, you may add a final concentration of 20% glycerol to the bacterial liquid. Please pay attention to the seal to avoid contamination. If the culture time is longer than 4h, it can be provided directly.
3)Bacteria grown on plate
The samples that need to be sequenced are grown on a plate by coating or line classification to form a single colony that can be seen by the naked eye. Please mark the number of colonies that need to be selected on the back of the plate. Please seal and pack carefully before sending for sequencing. If the number of colonies are not identified, we can get the monoclonal colony directly.
4) Glycerol bacteria
After freezing at -80 ℃, please resuscitation and provide it.
Plasmid extraction kit of QAIGEN, Promega, AXYGEN, Sangon and Takara are recommended to extract plasmids.
As far as possible, the OD value (260/280) of the tested plasmid should be guaranteed between 1.8 and 2.0. The plasmid concentration should be above 50ng/ul, and the volume should be above 10ul (single sequencing reaction).
Plasmids are preferably dissolved in sterile deionized water or in the eluent contained in the kit, rather than in TE.
If the amount of plasmid provided is too small, the cost of plasmid transformation by the company is required.
6) unpurified PCR products
Please provide more crude PCR products than 50ng/ul and 30ul above.The customer needs to provide the size of the Target band.
If the sample of the customer is not able to meet the requirement of delivery, we can arrange the purification experiment as far as possible according to the customer sample situation.
7) purify PCR products Please provide samples purified by gel purification or precipitation method, and electrophoresis detection shows that the band is single and bright, about 5-30ng/ul, and 10ul is dissolved in sterilized deionized water, rather than in TE.
For PCR samples whose length is less than 200bp or higher than 2000bp, it is suggested to be sequenced after cloning.If the PCR product shows multiple bands or dispersive samples, please re-prepare the sample or clone it and then provide the sample for sequencing.
The primers provided are best purified by PAGE, without the annexation bases, and are not specifically amplified. At the same time, the primer concentration should be marked. It is also possible to provide a sequence for synthesis.
1) Puncture bacteria