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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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4.8 ★★★★★
Based on 1066 reviews
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Product Reviews
★★★★★ 5
Interesting
Format: Kindle
So I will say I enjoyed the story, for sure had its moments where it dragged but it was a great story. I really liked that omegas picked their alphas/make the pack. Normally the Alphas make it and the omega fits in with them which is great but I enjoyed this new version where all the power basically went to the omega. It was a nice change of pace. I can admit some of the weird bedroom stuff with her being pregnant was odd, it’s really not hard to do stuff when pregnant (I know I’ve had two and it’s normal and even encouraged at the end especially if you want the baby out). But I like the story as a whole and will read the second, I do hope the next one isn’t dragged bc it stopped being action or tense after she met her alphas and I don’t think it was brought up or properly done when they tried to do it. More sweet after she left.
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Reviewed in the United States on November 11, 2024
★★★★★ 3
I’m a little disappointed.
Format: Kindle
I usually like Jillian West’s books but this one was missing a lot for me. The pregnancy didn’t come across as real. She’s on her feet for 12 hour days but is perfectly healthy at 8 months pregnant? Yet the week she moves in all of a sudden she’s not? She is planning on actually running during one of the plot buildups. But at 8 months pregnant that’s incredibly hard to do. The lack of breathing ability and lung space, the change in body center, mass, and gravity. All of it prohibits running, unless you’re an athlete this didn’t come off as at all realistic.
I didn’t feel any connection with the alphas. There wasn’t any emotional connection. It could be because of the tense it was written in. But I didn’t get any deep feelings out of this. It came across as checking off boxes. Even the spicy scenes weren’t really believable for me. I wanted to see them fall for her, and it just kind of all fizzled. Even Bishop.
One thing I did really like was the ending. I did not see it coming and I’m interested in reading book two because of it. But on the whole this book was mostly disappointing for me.
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Reviewed in the United States on March 16, 2024
★★★★★ 4
4.25 stars
Format: Kindle
Vale is an 8 month pregnant omega working as a waitress at a strip club and a cam girl. She starts to get very creepy vibes from a regular at the club, and her baby daddy ghosted her. She has had an online relationship with a man named Bishop through her cam girl status. One night, bishop was paying to watch her sleep and ansthe creepy regular Andrew break in and watch her sleep he tells vale to come to him at his business now. She flees and finds herself at a large security company with some.hot of alphas who are there to help her. This imegaverse is a little different than I have read, but I am thoroughly enjoying it. Vale is not a traditional omega she was raised by a single beta mom, and the alphas are not normal alphas they have never really loved pack life. But they are ruthless mercenaries. They need her, and she needs them. I love the aspect of the stalker and now the plot twists at the end, so so good. Sometimes, it seemed a little slow and stale mated, but since this a duet, I think It was just her starting to have Vale get to know her alpha suitors. Cliffhanger for sure with this one.
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Reviewed in the United States on September 9, 2024
★★★★★ 5
That ending 😫
Format: Kindle
I fell into a false sense of security and really thought this was gearing towards a happy ending. Then I realized there’s no work they don’t punish Andrew. I really liked Vale’s character. I don’t normally read books with pregnancy but going into this knowing she was pregnant made it more enjoyable for me.
I loved Bishops devotion to her and her happiness. I also loved that Holt and Mercy couldn’t fight their attraction to her. I love scent matches so very much. I’m so curious to see how this duet will end up. And I need to pay more attention and notice that a book I’m starting is a duet to begin with lol
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Reviewed in the United States on February 21, 2025
★★★★★ 5
oh wow
Format: Kindle
I just knew there was something about Cooper! I’m wondering if he’s about to be included but damn I’m glad he’s at least not a rapist and creepy guy, he just got called on assignment and had to go! This should be interesting! She’s gonna run and then what’s his face is gonna grab her. I’m worried! Wow that was a great book and cliffhanger! Loving this!
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Reviewed in the United States on December 27, 2025
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