今天的试验,plate孔内的1 N NaOH 100ul凭空消失,匪夷所思,百思不得其解

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实验步骤简单如下:

SOFIA with Rolling circle DNA amplification (T4 ligase normal standard)

1. Coat plate with : Anti-GFAP antibody.

2. block

3. serum(14samples)

4. Biotin-anti GFAP antibody.

5. Streptavidin

6. Primer: biotin-prostate-primer

7. T4 ligase

PEG4000

    phos-template

8.ф29 DNA polymerase buffer 

       BSA

      each DATP,DGTP,DCTP

       DTTP+DUTP-Texas Red 

      ф29 DNA polymerase

      PBS 

9. 1 N NaOH 

10. Laser test

今天的试验,神出鬼没,试验plate孔内的1 N NaOH 100ul凭空消失,匪夷所思,百思不得其解。

Plate 1

Plate 1, C 和E的3,4,5,6,7,8,9,是样品1,2,3,4,5,6,7,C和E的1,2,10,11,12是PBS。这里最重要的条件是:C和E完全一样。

plate 2

Plate 2, C 和E的3,4,5,6,7,8,9,是样品8,9,10,11,12,13,14,C和E的1,2,10,11,12是PBS。这里最重要的条件是:C和E完全一样。

实验进行到第9步骤,就是加了1 N NaOH (100ul ),零下20度过夜。第二天要进行Laser test的时候,发现,

1. Plate 1, C的3,4,5,6,7,8,9内的100ul 1 N NaOH,不见了,而与之对应的Plate 1, E的3,4,5,6,7,8,9内的100ul 1 N NaOH还在,而C和E的1,2,10,11,12的内的100ul 1 N NaOH还在。

2. Plate 2, C的3,4,5,6,7,8,9内的100ul 1 N NaOH,不见了,而与之对应的Plate 2, E的3,4,5,6,7,8,9内的100ul 1 N NaOH还在,而C和E的1,2,10,11,12的内的100ul 1 N NaOH还在。

这消失的100ul 1 N NaOH,去了哪里?

假如是serum的区别,就是C和E的1,2,10,11,12的内的100ul 1 N NaOH还在(就是PBS还在,而serum不在了),那么,为什么, E的3,4,5,6,7,8,9内的100ul 1 N NaOH还在?而C的3,4,5,6,7,8,9内的100ul 1 N NaOH消失了呢?

这里一个重要的信息是,剩余的 1 N NaOH几乎是100ul,而消失的1 N NaOH的plate孔,很干净,几乎看不见任何液体。

这凭空消失的条件是什么?关键是与之完全对应的E的3,4,5,6,7,8,9内的100ul 1 N NaOH还在.

 

 

 

 

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