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Description
Organotal rat normal small intestinal organoid culture KitProduct Specification Usage 1. Primary (1) the normal small intestine tissues of rats after sampling must be cleaned at 2 8 , several Petri dishes should be prepared, and primary culture buffer B pre cooled at 4 should be added for standby (double antibody and gentamicin should be added) (2) put the tissue into the Petri dish, wash it three times with primary culture buffer B, remove impurities, remove the intestinal mucosa of normal small intestine
Product Specification
| Usage |
1. Primary (1) the normal small intestine tissues of rats after sampling must be cleaned at 2-8 ℃, several Petri dishes should be prepared, and primary culture buffer B pre cooled at 4 ℃ should be added for standby (double antibody and gentamicin should be added) (2) put the tissue into the Petri dish, wash it three times with primary culture buffer B, remove impurities, remove the intestinal mucosa of normal small intestine tissue of rats with ophthalmic scissors or scalpel, and cut it into a tissue block with a volume of about 10-30mm3 (3) the tissue was digested with rat normal small intestine primary tissue digestive solution C, and then shaken at 4 ℃ for 20-25min or left to digest for 40-50min (observe the digestion at any time during the digestion process) (4) take a small amount of liquid and observe under the microscope. More crypts are observed under the microscope. Add three times the volume of primary culture buffer B to stop digestion (5) filter with a 100um pore size screen, collect the filtrate, remove the supernatant after centrifugation at 300g for 5min, add primary culture buffer B and resuspend for centrifugation (6) Matrigel calculation: after step 5, observe the collected tissue volume, add 25 times the tissue volume of Matrigel (abs9495) to resuspend the plate (7) take a 24 well cell culture plate as an example, dispense 25ul of tissue Matrigel mixture per well for plating (operate at 4 ℃) (8) put the laid culture plate into a 37 ℃ incubator for 10-15min to form a glue, and add rat normal small intestinal organoid medium a (restore room temperature) for culture 2. Organoid subculture (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) a: the number of organoids is insufficient or the volume is small: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4 b: When the number of organoids is large or the volume is large: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid passage digest D for digestion for 2-3min, add organoid passage culture buffer g to stop digestion, centrifuge for 5min to discard the mixture, add an appropriate amount of organoid passage culture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4 (4) after organoids were collected, add Matrigel to resuspend, lay 25ul Matrigel per well in a 24 well cell culture plate, place in the incubator for 10-15min, and add 500ul of rat normal small intestinal organoid culture medium a 3. Organoid cryopreservation (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend, and centrifuge for 5min at 300g to discard the liquid (4) add an appropriate amount of organoid cryopreservation solution F, gently blow and resuspend. Take a 24 well cell culture plate as an example: the density is 2 wells and 1 tube is frozen, with a volume of 1.4ml per tube (5) mark the information, and after cooling down the program, transfer it to liquid nitrogen for long-term storage 4. Organoid resuscitation (1) take 10ml of organoid subculture buffer g and put it into a 15ml centrifuge tube (2) take out the frozen organoid cells from the liquid nitrogen tank and quickly put them into a 37 ℃ water bath pot for melting (3) during the water bath thawing process, it is necessary to gently shake the freezing tube to ensure that the frozen solution is completely thawed in 1-2min (4) quickly transfer the dissolved organoid cells to a 15ml centrifuge tube, gently blow 6-8 times with a pipette, centrifuge at 300g for 5min, and then remove the supernatant and collect the organoid cell pellet. Add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube. Centrifuge for 5min at 300g (5) resuspend Matrigel, lay 25ul Matrigel per well in a 24 well cell culture plate, place in an incubator for 10-15min to form a gel, and add 500ul of rat normal small intestinal organoid culture medium A. |
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| Synonym | Rat Small Intestine Organoid Culture Kit | ||||||||||||||||
| Description | Composition:
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| Storage Temp. | 4 ℃, 3months; -20 ℃, 1 year, see the label for details |
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4.2 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 5
A Beautiful Timepiece at an Awesome Price!!
Color: Grey, Color: Grey
I have lusted after this watch for four months. I first tried on a similar model (stainless steel band with a green face) in a local jewelry shop (Spitz Jewlers, Walnut Creek, CA, USA). It was "love at first sight," even if I didn't love the white lines on the face from 12-6 and 3-9. At the time, the MSRP was $775.
The MSRP has since increased to $795. But I bought this beauty on Amazon.com for only $524.99; after applying the cash-back from my Prime credit card, this watch costs only $498.75. This was "the offer that I could not refuse," to quote from "The Godfather."
Accuracy could be better: after only a few days, it is now 7 seconds behind my phone (to which I set my new watch). But I don't really care about that all that much. This is why I rated "accuracy" at 4 stars, but my overall review is 5 stars.
My video shows the date changing suddenly at 11:59:59pm, just how I want it; in another photo, you can see that my Seiko changes the date more slowly, which I do not like.
This watch is absolutely phenomenally beautiful. If you're on the fence over whether to make this purchase, then I would say "go for it!" You will not find a prettier, more accurate watch at a lower price in ANY store in the entire USA.
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Reviewed in the United States on April 28, 2023
★★★★★ 4
Classy and sporty. Great value!
Color: Grey, Color: Grey
Excellent watch for the price. Looks sporty and dressy. More towards dress watch. Blue is dynamic. Strongly recommend blue for sporty feel. I was thinking of aqua terra. This come close to it with a price less than what i would have paid my state tax for omega aqua terra. My only concern is bracelet which i feel kind of OK. Better bracelet design would have made this watch much superior. 21 lug width is awful. Other than that, nothing to complain.
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Reviewed in the United States on July 13, 2020
★★★★★ 5
Stunning Gentleman's Watch
Color: Grey
This is my second Tissot watch. It's quite stunning in person. The online photos don't do it justice. It has the look and feel of a much more expensive gentleman's watch which makes this purchase a great bargain. Tissot is a great reliable Swiss made watch brand. My first Tissot watch which is probably more than 20 years old still runs great. Very pleased with this purchase.
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Reviewed in the United States on October 16, 2025
★★★★★ 5
Good Tissot Watch
Color: Grey
Arrived on time, in its original box, include a 2 year international warranty, into the box include a book with the Tissot brand history in Switzerland. Algo a second book with a whole catalog of Tissot watches men and women.
The watch looks better when you put in on your wrist. The “naval” blue really gives the clock a classy look. The stainless steel workmanship is really good, the metal is bright and perfectly polished. The metal wristband quality is very good, did not have any problem to adjust the size , I went to my preferred clock workshop for this job, they extracted a few pieces of the band to fit to my small wrist size. Have to get used to the butterfly latch, but after a few days, now I learned to open and close it without any effort. The clock it’s very accurate, I’ve been using my Tissot for near a month and it just increased a few seconds per week. I do the setup and check accuracy with my pc clock. In short, It”s a good value watch, nice, good looking, accurate.
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Reviewed in the United States on August 1, 2021
★★★★★ 5
Durable
Style: Bi-fold
My man loves it. He works construction and it has held up well so far. Carhartt does good work. Looks exactly like the picture, well made.
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Reviewed in the United States on April 19, 2026
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