Milk Creamatocrit - Revision history

Mollyec at 20:29, 18 March 2022

2022-03-18T20:29:59Z

← Older revision		Revision as of 20:29, 18 March 2022
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	==Mouse Milk Creamatocrit (milk fat %)==		==Mouse Milk Creamatocrit (milk fat %)==
			This method is done to assess the total fat content of mouse milk. It does not differentiate between type of fat present. Additional information on mouse milk composition can be ascertained through triglyceride or cholesterol assay, or through lipidomics.
	===Materials===		===Materials===
	*Milk samples (from -80)		*Milk samples (from -80)
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	* After the 8th spin, immediately read the creamatocrit using the calipers		* After the 8th spin, immediately read the creamatocrit using the calipers
	===Reading the samples===		===Reading the samples===
	* Measure to the nearest 0.01 inch the aqueous layer, the fat layer, and the small aqueous layer on top of the fat layer.		* Measure each of the 3 layers to the nearest 0.01 inch with the calipers. Record the number in the corresponding columns of [https://docs.google.com/spreadsheets/d/e/2PACX-1vTm1Zo5Cf9RTuw_uRwum8h_dOBb0mjc5HYcOHR1nBPHKbSzxlFqwYfJs0NMdChFHrczE_8sKCU3K9lp/pub?output=csv the spreadsheet](the aqueous layer(Aqueous.inches.1), the fat layer(Fat.inches), and the small aqueous layer on top of the fat layer(Aqueous.inches.2)).
	* Record results in a csv file.
	* Calculate the milk fat percentage using the following R code:		* Calculate the milk fat percentage using the following R code:

Davebrid: /* Reading the samples */

2022-03-17T19:04:12Z

Reading the samples

← Older revision		Revision as of 19:04, 17 March 2022
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	* Calculate the milk fat percentage using the following R code:		* Calculate the milk fat percentage using the following R code:

	```{r data-manipulation}		```{r data-manipulation}
	mutated.milk.data<-milk.data%>%		mutated.milk.data<-milk.data%>%
	group_by(replicate, MouseID, Genotype)%>%		group_by(replicate, MouseID, Genotype)%>%
	mutate(fat = Fat.inches*25.4,		mutate(fat = Fat.inches*25.4,
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	mutate(total.volume = water.corrected+fat)%>%		mutate(total.volume = water.corrected+fat)%>%
	mutate(fat.percent = (fat/total.volume)*100)		mutate(fat.percent = (fat/total.volume)*100)
	```		```

Mollyec: /* Reading the samples */

2022-03-17T15:44:23Z

Reading the samples

← Older revision		Revision as of 15:44, 17 March 2022
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	* Calculate the milk fat percentage using the following R code:		* Calculate the milk fat percentage using the following R code:

			```{r data-manipulation}
	mutated.milk.data<-milk.data%>%		mutated.milk.data<-milk.data%>%
	group_by(replicate, MouseID, Genotype)%>%		group_by(replicate, MouseID, Genotype)%>%
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	mutate(total.volume = water.corrected+fat)%>%		mutate(total.volume = water.corrected+fat)%>%
	mutate(fat.percent = (fat/total.volume)*100)		mutate(fat.percent = (fat/total.volume)*100)
			```

Mollyec: added more to materials

2022-03-17T15:43:00Z

added more to materials

← Older revision		Revision as of 15:43, 17 March 2022
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	==Mouse Milk Creamatocrit (milk fat %)==		==Mouse Milk Creamatocrit (milk fat %)==
	===Materials===		===Materials===
			*Milk samples (from -80)
			*Ice bucket
	*CritSpin Micro-Hematocrit Spinner (from the Pennathur lab in Brehm tower)		*CritSpin Micro-Hematocrit Spinner (from the Pennathur lab in Brehm tower)
	*1XPBS + EDTA (50mL 1X PBS + 100uL 0.5M EDTA)		*1XPBS + EDTA (50mL 1X PBS + 100uL 0.5M EDTA)
	*~~untreated~~ glass hematocrit tubes		*Untreated glass hematocrit tubes
	*Fractional dial calipers		*Fractional dial calipers
	*Critoseal		*Critoseal

Mollyec: generated protocol from Brigid's creamatocrit SOP

2022-03-15T13:57:18Z

generated protocol from Brigid's creamatocrit SOP

New page

==Mouse Milk Creamatocrit (milk fat %)==
===Materials===
*CritSpin Micro-Hematocrit Spinner (from the Pennathur lab in Brehm tower)
*1XPBS + EDTA (50mL 1X PBS + 100uL 0.5M EDTA)
*untreated glass hematocrit tubes
*Fractional dial calipers
*Critoseal
*Kimwipes
*P10 pipette with tips

===Preparing the samples===
#Thaw whole milk on ice. To homogenize thawed milk, pipette up and down for five seconds. DO NOT shake!
#Dilute the milk in aa 1:3 solution with the 1XPBS + EDTA. Make enough for 4 capillary tubes
#Take capillary tube and draw in enough milk to fill ~3/4 of the tube (stop before the blue line)
#Hold horizontally and seal the other end of the tube with critoseal twice. Be sure to continue to hold the tube horizontally, or the milk will be ejected.
#Wipe excess milk from the tube with the kimwipe
#Each sample should be run in duplicate (you may need to run additional tubes if the variance between the first 2 sample results is >=25%)
===Running samples===
* Place sealed and filled capillaries into the critspin micro-hematocrit spinner. The sealed end should point toward the outer edge of the centrifuge.
* Record the position of each tube/sample before running..
* Screw on the centrifuge cover so that it is secure, but not too difficult to loosen later.
* Spin for 120 seconds, eight separate times. Between spins, the centrifuge will come to a complete stop on its own.
* After the 8th spin, immediately read the creamatocrit using the calipers
===Reading the samples===
* Measure to the nearest 0.01 inch the aqueous layer, the fat layer, and the small aqueous layer on top of the fat layer.
* Record results in a csv file.
* Calculate the milk fat percentage using the following R code:

mutated.milk.data<-milk.data%>%
group_by(replicate, MouseID, Genotype)%>%
mutate(fat = Fat.inches*25.4,
water = (Aqueous.inches.1 + Aqueous.inches.2)*25.4)%>%
mutate(water.corrected = water/4)%>%
mutate(total.volume = water.corrected+fat)%>%
mutate(fat.percent = (fat/total.volume)*100)