"
Team:Cambridge-JIC/Guide/Care/SexualReproduction
From 2014.igem.org
(Created page with "The sexual phase can be induced by incubating Marchantia in far red light (650nm+) for approximately 2 weeks. 1. Pipette ~ 50-100 ul sterile water onto sexual organs 2. Using a ...") |
|||
| Line 1: | Line 1: | ||
| + | =Sexual Reproduction= | ||
| + | You will need to do this to collect spores for transformation. See it as 'making a competent stock'. The following protocols apply for working with Marchantia growing in soil, or in sterile culture. | ||
| + | |||
| + | ==Inducing sexual reproduction== | ||
The sexual phase can be induced by incubating Marchantia in far red light (650nm+) for approximately 2 weeks. | The sexual phase can be induced by incubating Marchantia in far red light (650nm+) for approximately 2 weeks. | ||
| + | ==Fertilising== | ||
1. Pipette ~ 50-100 ul sterile water onto sexual organs | 1. Pipette ~ 50-100 ul sterile water onto sexual organs | ||
2. Using a sterile brush, brush spermatozoa from archaegoniaphore onto female sexual organ | 2. Using a sterile brush, brush spermatozoa from archaegoniaphore onto female sexual organ | ||
3. Sporeheads start to be visible in 2 weeks | 3. Sporeheads start to be visible in 2 weeks | ||
| + | |||
| + | ==Storing sporangia== | ||
| + | Cut individual sporeheads and store with silica beads at -80 for long term storage, or at -20 for medium term storage. For short term use, sporangia can be left on a bench indefinitely, with little loss of viability. | ||
| + | |||
| + | ==Notes== | ||
| + | Try and be sterile! Fungal contamination is persistent and difficult to get rid of. | ||
Revision as of 01:02, 17 October 2014
Contents |
Sexual Reproduction
You will need to do this to collect spores for transformation. See it as 'making a competent stock'. The following protocols apply for working with Marchantia growing in soil, or in sterile culture.
Inducing sexual reproduction
The sexual phase can be induced by incubating Marchantia in far red light (650nm+) for approximately 2 weeks.
Fertilising
1. Pipette ~ 50-100 ul sterile water onto sexual organs 2. Using a sterile brush, brush spermatozoa from archaegoniaphore onto female sexual organ 3. Sporeheads start to be visible in 2 weeks
Storing sporangia
Cut individual sporeheads and store with silica beads at -80 for long term storage, or at -20 for medium term storage. For short term use, sporangia can be left on a bench indefinitely, with little loss of viability.
Notes
Try and be sterile! Fungal contamination is persistent and difficult to get rid of.
