試管培養(yǎng)基上有冷凝水怎么處理？
發(fā)布時(shí)間：
2022-12-24
作者：
無(wú)菌培養(yǎng)基/平板培養(yǎng)基
很多朋友在制作瓊脂母種培養(yǎng)基的時(shí)候，在滅菌冷卻后的母種培養(yǎng)基上往往有很多冷凝水，擔(dān)心對(duì)接種后的菌種有不良影響。冷凝水影響菌種的生長(zhǎng)嗎，答案是肯定的，特別是在接種金針菇、蟹味菇、黃傘、蟲(chóng)草等菌種時(shí)，如果母種培養(yǎng)基帶有比較多的冷凝水，那么以上這幾個(gè)品種的菌絲生長(zhǎng)都不會(huì)太好看。怎么可以避免這種情況，如何才能做得更好，菇園彩風(fēng)給大伙提幾點(diǎn)建議，以供參考。
一、培養(yǎng)基滅菌計(jì)時(shí)結(jié)束，壓力表指針回零后，可以將高壓鍋鍋蓋打開(kāi)，但不拿走，在鍋上移開(kāi)一道縫隙，讓鍋內(nèi)的熱氣涌出，讓棉塞和試管內(nèi)的水蒸氣先蒸發(fā)一部分，約10分鐘后，再將母種培養(yǎng)基取出，這對(duì)預(yù)防冷凝水的形成有很大的作用；
二、母種培養(yǎng)基滅菌完畢，取出擺放斜面后，可以用多層厚厚的干凈紗布等將剛擺好的試管培養(yǎng)基蓋好，一是可以防止在冷卻過(guò)程中，有灰塵降落在試管培養(yǎng)基表面，影響以后的無(wú)菌操作；二是可以防止在冷卻過(guò)程中，玻璃試管內(nèi)外溫差過(guò)大，盡量降低冷凝水的形成。實(shí)際這個(gè)方法還不能完全杜絕冷凝水的形成；
三、如果使用硅膠塞封試管口，可提前一周將母種培養(yǎng)基做好，如果用棉塞封口，時(shí)間可以適當(dāng)縮短，這樣經(jīng)過(guò)一周左右的靜置，母種培養(yǎng)基管壁和斜面上的冷凝水可以基本蒸發(fā)掉，在使用時(shí)就會(huì)非常方便。這一點(diǎn)可能有很多人反對(duì)，因?yàn)楹芏嘟炭茣?shū)上說(shuō)，母種培養(yǎng)基要現(xiàn)做現(xiàn)用，或盡快使用，否則容易被雜菌污染，菇園彩風(fēng)認(rèn)為沒(méi)有那么嚴(yán)重，只要母種培養(yǎng)基滅菌徹底，放置時(shí)間再長(zhǎng)也沒(méi)有什么問(wèn)題，這一點(diǎn)大可放心。不過(guò)在放置時(shí)要注意環(huán)境衛(wèi)生，培養(yǎng)基試管口要稍微抬起放置，不能放在灰塵很多的地方，試管口可以用牛皮紙包好，特別是棉塞封口的培養(yǎng)基不能密封保存，防止棉塞滋生雜菌，這一點(diǎn)很重要。四、如果實(shí)在要急用培養(yǎng)基，帶冷凝水接種也可以，有可能在菌絲生長(zhǎng)時(shí)，會(huì)受到冷凝水的影響，生長(zhǎng)暫時(shí)可能表現(xiàn)不良，不過(guò)沒(méi)有問(wèn)題，隨著水份的蒸發(fā)和菌絲的快速生長(zhǎng)，慢慢就會(huì)表現(xiàn)正常。這里提幾個(gè)細(xì)節(jié)，如果帶水接種，在母種培養(yǎng)階段，母種培養(yǎng)基的口部要稍微抬起，防止冷凝水流到管口，打濕棉塞或硅膠塞，這個(gè)會(huì)對(duì)菌種的質(zhì)量產(chǎn)生不良影響；如果冷凝水太多，可以在接種時(shí)倒掉，這時(shí)要注意無(wú)菌操作，不能讓冷凝水回流，倒完后，繼續(xù)保持傾斜姿勢(shì)，在酒精燈火焰附近將管口灼燒后，才能開(kāi)始接種操作，嚴(yán)防管口帶菌，隨水回流進(jìn)入管內(nèi)，造成不必要的污染；也可以將管口先灼燒滅菌后，再將水倒掉。
When making agar-agar mother culture medium, many friends often have a lot of condensation water on the sterilized and cooled mother culture medium, worrying about the adverse effect on the inoculated bacteria. Condensate affect the growth of bacteria, the answer is yes, especially in the vaccination needle mushroom, crab flavor mushroom, yellow umbrella, such as Chinese caterpillar fungus strains, if mother with more kinds of culture medium condensate, so more than the several varieties of hypha growth is not too good. How can avoid this kind of situation, how can ability do better, mushroom garden color wind gives everybody a few Suggestions, for reference.
A sterilization time end, medium, pressure gauge pointer back to zero, you can open the pressure cooker pot, but does not take away, in the pan. Remove a gap, let the pot hot air flow, let the tampon and water vapor in vitro evaporate, about 10 minutes, then remove the mother kind of medium, which has great effect to prevent the formation of condensation water;
Second, the mother of medium after sterilization, after putting the cant take out, can with thick layers of clean cloth etc will build good, just set the in vitro culture medium can be prevented in the cooling process, the in vitro culture medium surface with ash landing, influence after aseptic operation; Second, it can prevent the temperature difference between the inside and outside of the glass tube from being too large in the cooling process, and minimize the formation of condensate water. In fact, this method can not completely eliminate the formation of condensate water;
Three, if you use silica gel plug seal tube mouth, mother kind of medium can be a week in advance, if the plug sealing cotton, time can be shorten appropriately, so after a week or so of rest, mother of medium tube wall and the condensed water can evaporate, basic on the slope when use is very convenient. That there may be many people opposed, because many textbooks say, mother kind of medium to use now now, or to use as soon as possible, otherwise easy to bacterial contamination, mushroom garden colorful wind don't think so serious, as long as the mother of medium sterilization thoroughly, place is no longer problem, it can rest assured. But should pay attention to the environmental health, when placed culture medium in vitro mouth slightly lift the place, not in the dusty, tube can be wrapped in kraft paper, especially the cotton plug sealing medium can't seal save, prevent cotton breeding of mixed bacteria, it is very important.
Fourth, urgent need medium, if at all, with condensed water inoculation can also, when hypha growth likely, are influenced by the condensed water, grow poorly may temporarily, but there is no problem, with the rapid growth of water evaporation and hyphae, slowly will appear normal. Here to ask a few details, if with water, in the training phase, the mother of mother culture medium of the mouth slightly raised, prevent the cooling water flow to the nozzle, wet cotton or silica gel seal, this will have bad effect to the quality of the strains; If the condensed water is too much, can pour out when vaccination, then pay attention to the aseptic operation, can't let condensate return, after the fall, continue to maintain the tilted position, near the alcohol lamp flame tube mouth after burning, can begin vaccination operation, keep a nozzle carrier, with the water back into the tube, the unnecessary pollution; Or you can burn the nozzle first to sterilize it and then pour out the water.