"
Restricting Plasmids (Double Restriction)
From 2008.igem.org
(Difference between revisions)
Riachalder (Talk | contribs) |
|||
| (One intermediate revision not shown) | |||
| Line 1: | Line 1: | ||
| + | {{:Team:Newcastle University/Header}} | ||
| + | {{:Team:Newcastle University/Template:UnderTheProtocol|page-title=[[Restricting Plasmids (Double Restriction)]]}} | ||
| + | |||
[[Image:Jess1.jpg|thumb|right|200px| Jess enjoying pipetting buffer into her restriction mixture.]] | [[Image:Jess1.jpg|thumb|right|200px| Jess enjoying pipetting buffer into her restriction mixture.]] | ||
We have conducted restrictions in varying concentrations and total volumes; however they all follow the same basic procedure. Below are two of the different restrictions we carried out. | We have conducted restrictions in varying concentrations and total volumes; however they all follow the same basic procedure. Below are two of the different restrictions we carried out. | ||
| - | |||
| Line 19: | Line 21: | ||
Total volume = 30μl | Total volume = 30μl | ||
| - | |||
| - | |||
| - | |||
| + | [[Image:DSC01597.jpg|200px]] | ||
| + | The DNA purification kit we use to purify enzymatic reaction mixtures. | ||
| - | + | * Incubate solutions for 90 minutes in a 37°C water bath. | |
| - | + | * If DNA is to be run on a gel and a band cut out, this can be done without purifying the DNA. For all other downstream applications or for storage, solution must be purified to remove the enzymes. | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
Latest revision as of 16:01, 28 October 2008
Newcastle University
GOLD MEDAL WINNER 2008
| Home | Team | Original Aims | Software | Modelling | Proof of Concept Brick | Wet Lab | Conclusions |
|---|
Home >> Wet Lab >> Protocols >> Restricting Plasmids (Double Restriction)
We have conducted restrictions in varying concentrations and total volumes; however they all follow the same basic procedure. Below are two of the different restrictions we carried out.
- 46μl MillQ H2O
- 10μl 10 x buffer
- 40μl plasmid sample
- 2μl enzyme 1
- 2μl enzyme 2
Total volume = 100μl
Concentrated
- 10μl MilliQ H2O
- 3μl 10 X buffer
- 10μl plasmid sample
- 1μl enzyme 1
- 1μl enzyme 2
Total volume = 30μl
The DNA purification kit we use to purify enzymatic reaction mixtures.
- Incubate solutions for 90 minutes in a 37°C water bath.
- If DNA is to be run on a gel and a band cut out, this can be done without purifying the DNA. For all other downstream applications or for storage, solution must be purified to remove the enzymes.
