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Table 4 Ability of each kit to effectively remove DNA from small tissue samples

From: A comparison of commercially-available automated and manual extraction kits for the isolation of total RNA from small tissue samples

 

Pre-DNase

Post-DNase

 

no RT

RT

Difference

no RT

RT

Difference

Plus Mini

      

Blood

33.65

31.71

1.94

> 40*

32.23

7.77

Spleen

39.84*

28.62

11.22

39.84*

29.59

10.25

Kidney

38.06*

19.35

18.71

> 40*

23.41

16.59

Embryo

> 40*

27.82

12.18

> 40*

29.46

10.54

Larvae

> 40*

26.26

13.74

> 40*

27.86

12.14

Plus Universal

      

Blood

33.27

32.87

0.40

> 40*

34.07

5.93

Spleen

34.77

28.90

5.87

39.92*

29.14

10.78

Kidney

36.50*

21.54

14.96

> 40*

22.18

17.82

Embryo

39.79*

29.23

10.56

> 40*

31.17

8.83

Larvae

> 40*

20.74

19.26

> 40*

28.79

11.21

Maxwell®

      

Blood

> 40*

34.19

5.81

> 40*

34.84

5.16

Spleen

> 40*

29.38

10.62

> 40*

30.25

9.75

Kidney

> 40*

20.52

19.48

> 40*

20.67

19.33

Embryo

37.09*

26.41

10.68

> 40*

25.86

14.14

Larvae

> 40*

26.46

13.54

> 40*

27.55

12.45

MagMAX ®

      

Blood

29.79

28.11

1.68

39.02*

32.65

6.37

Spleen

> 40*

29.93

10.07

38.04*

30.36

7.68

Kidney

31.83

21.89

9.94

39.74*

21.51

18.23

Embryo

36.34*

22.57

13.77

> 40*

22.31

17.69

Larvae

33.75

27.54

6.21

> 40*

28.17

11.83

SimplyRNA HT

      

Blood

> 40*

31.06

8.94

> 40*

31.55

8.45

Spleen

> 40*

28.11

11.89

> 40*

27.9

12.10

Kidney

> 40*

21.54

18.46

> 40*

21.94

18.06

Embryoa

-

-

-

-

-

-

Larvae

> 40*

26.22

13.78

> 40*

26.00

14.00

  1. The threshold cycle (Ct) values of samples subjected to the kit DNase treatment only (pre-DNase) or to an additional DNase treatment, as well as the corresponding "no RT" samples (see Methods for details). *represents no RT samples with Ct values ≥35 and thus, considered gDNA free. indicates samples for which difference between the Ct values of the no RT and RT samples was ≥5, an indicator of sufficient gDNA removal for qPCR gene expression analysis.
  2. aThe quantities of RNA isolated from embryos using the SimplyRNA HT kit were less than the amount necessary for the first-strand synthesis protocol used in this study; therefore, qPCR reactions could not be performed with these samples.