如何制作灵芝母种培养基?

2025-03-16 04:11:41
推荐回答(1个)
回答1:

(1)称取

接配方精确称取各种营养物质。

(2)制备

马铃薯等块茎、植物根去皮后切块洗净,加水1000毫升,文火煮沸30分钟后过滤取汁;玉米粉称重后加水1000毫升,加热到70℃左右,保持60分钟,过滤取汁;麦芽称重后加水1000毫升,加热至60℃左右,保持60分钟,过滤取汁,最后补足水量至1000毫升。

(3)酸碱度调整

灵芝一般要求pH5.5~6,用稀碱1摩尔/升NaOH、稀酸1摩尔/升HCl来调整酸碱度至适度为止。

(4)分装、灭菌存放

将称好的琼脂,加入上述配好的液体培养基中,边加热边搅拌,待琼脂完全熔化后,用湿纱布迅速过滤,趁热进行分装。分装前要备好容器,分装试管时,要用玻璃漏斗进行分装,尽量避免培养基黏附管口,如培养基不慎黏着管口或管壁,要用纱布擦净,以免培养基粘住棉塞而引起污染。试管装量视需要而定,斜面培养基,装量为试管长度的1/5或1/4;深层培养基,一般为试管长度的1/2或2/3。培养基分装好后均要加上棉塞。棉塞用普通棉花制作,不宜使用脱脂棉。棉塞的大小,松紧要与试管口的口径相称,塞入试管的棉塞要紧贴管壁,不留缝隙,松紧度以提起棉塞而试管不脱落,拔出棉塞有轻微的声音为适宜。标准的棉塞应是塞头略大,不易变形,入管的部分占塞总长的2/3,外露1/3为宜;然后把2~3只试管捆扎在一起,棉塞外包扎牛皮纸或旧报纸。再用油纸包扎,装入高压灭菌锅灭菌,装锅时试管均要竖放,切勿倾斜或平置。蒸汽压力1.2千克/厘米2,温度达121℃,保持30分钟。灭菌后,让其压力和温度自然下降之后,取出培养基按要求摆成斜面,斜面的长度以不超过试管的1/2为宜,培养基冷却凝固,即成斜面培养基。放在25~28℃条件下,空白培养1~2天,检查无污染,才能使用。

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