实验动物模型

Purpose: To perform restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion. Time required: 2-4 hours Special Reagents: dependent upo ...

Pick colonies into 0.5ml of SD-Leu (or other appropriate SD medium) Vortex for 1min Leave to grow O/N for 18-24h at 30°C 230-250rpm (best in 5ml bijou) Spin yeast culture at 13000rpm for 5 min (microf ...

Grow a 5 ml YPD O/N culture inoculated with a single yeast colony at 30 deg. Transfer culture to a small 13 x 100 glass tube. Spin down cells 2 min. in tabletop centrifuge. Pour off supernatant. Wash ...

Grow 100 ml yeast cells in desired media overnight to an A600 of ~1.0 (0.6 to 1.2 works well). For growth in minimal media 1 ml of a saturated overnight culture in minimal media (Synthetic dextrose (S ...

Yeast Media: Note: Synthetic complete medium can be prepared by adding media supplements (see below). Medium using 6.7 g yeast nitrogen base without amino acids (Difco # 0919-15) can also be prepared ...

Cells are grown for 2-3 days as 1.5ml prep. under selection for the plasmid of interest. Spin cells down 2.6K for 5min. Resuspend in 1ml 0.25m NaOH/1% 2-mercaptoethanol Incubate 10min on ice. Add 0.16 ...

-Use sterile technique and sterile solutions in steps 1 to 3.- 1. Using a saturated starter culture inoculate 25 to 30 ml of appropriate media in a 125 ml flask. ***Since it is often difficult to esti ...

Procedure pick one colony inoculate in 3 ml of the appropriate media grow at 30° overnight pellet the cells (5 min 5000g) wash 1X in sterile ddH2O re- suspend the pellet in 200 µl EthOH (optional: +2 ...

This is the preferred method for yeast RNA preparation use Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fresh YPD media and grow cells at 30 degrees overnigh ...

Day 1Set up an overnight culture in 100 ml LMM broth or 100 ml terrific broth containing 100ul 100 mg/mlAmp Day 2Add 40-50 ml o/n culture to 1 lt terrific/K2K (see appendix) with 0.5 ml 100mg/ml Amp. ...

Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically add 0.1 to 0.2 ml saturated culture in the evening to get an of OD600 = 1 to 2 the next morning. ...

Transformation Protocol for Arabidopsis � Abbreviated Germinate seed in pots ↓ 4 weeks Streak bacteria onto YM/MinA ↓ 2-3 days 28°C Spray/dip bacterial suspension onto plants ↓ 1 day in box Let plants ...

This is a simple and fast protocol for the extraction of genomic DNA from Arabidopsis thaliana that works fine in PCR for simple amplicons. We only use this with (rosette) leaves but it is likely that ...

OverviewIn order to study spleen cells (e.g. lymphocytes granulocytes other immune cells) it helps to make single-cell suspensions so that the cells can be manipulated ex vivo easily. This protocol su ...

One of the uses of tamoxifen in the lab is the activation of enzymes deactivated via an estrogen-binding domain like Cre-ERT (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinit ...

PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al. Methods Enzymol. 254:3-20 (1995). The epidermis from a newborn mou ...

Setup You have yeast strains that are deficient in mating (eg Ste12 knockouts) and would like to test whether transforming them with a plasmid that contains genes that are supposed to complement the m ...

MethodUsing sterile pipette tips transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase 3 uL of 1 U/mL Zymolyase stock solution 47 uL of water Incubate cell suspension at 37 C for 30 minutes. Incubate ...

Materials (Solutions are all available from the media room) 200ml bottle of 2x SD 200ml bottle of 4% agar -- make sure to sign it out 40% glucose CSM minus the relevant marker(s) as powder Stack of pl ...

General Information The cells being plated are sandwiched between layers of cell-free agar. The submerged cells grow into smaller colonies and many more colonies can be counted per plate (I''ve been a ...